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Abstract:

The present disclosure provides topical biophotonic materials and methods
useful in phototherapy. In particular, the topical biophotonic materials
of the present disclosure include a cohesive matrix, and at least
chromophore which can absorb and emit light from within the topical
biphotonic material, wherein the topical biophotonic material is elastic.
The topical biophotonic materials and the methods of the present
disclosure are useful for promoting wound healing and skin rejuvenation,
as well as treating acne and various other skin disorders.

Claims:

1. A topical biophotonic material comprising: a cohesive matrix, and at
least one chromophore which can absorb and emit light from within the
biophotonic material, wherein the topical biophotonic material is
elastic.

4. The topical biophotonic material of claim 1, wherein the tear and/or
tensile strength of the biophotonic material is greater than an adhesive
strength of the topical biophotonic material to a surface to which it is
applied.

10. The topical biophotonic material of claim 9, wherein the pre-formed
configuration is a shape and/or a size corresponding with a shape and/or
a size of a body part to which the topical biophotonic material can be
applied.

12. The topical biophotonic material of claim 9, wherein the topical
biophotonic material is a mask.

13. (canceled)

14. The topical biophotonic material of claim 12, wherein the mask is a
face mask having at least one opening for the eyes, nose or mouth.

15. The topical biophotonic material of claim 9, wherein the preformed
configuration is a shape and/or a size corresponding with a shape and/or
a size of a light source or lamp to which the topical biophotonic
material can be attached.

16. The topical biophotonic material of claim 1, wherein the topical
biophotonic material can be removed without leaving substantially any
residue on a surface to which the topical biophotonic material is
applied.

17-19. (canceled)

20. The topical biophotonic material of claim 1, wherein the chromophore
is a xanthene dye.

66. A method for biophotonic treatment of a skin disorder comprising:
placing a topical biophotonic material over a target skin tissue, wherein
the topical biophotonic material is elastic and comprises at least one
chromophore and a cohesive matrix; and illuminating said topical
biophotonic material with light having a wavelength that overlaps with an
absorption spectrum of the at least one chromophore; wherein said
biophotonic material emits fluorescence at a wavelength and intensity
that promotes healing of said skin disorder.

67-68. (canceled)

69. A method for biophotonic treatment of acne comprising: placing a
topical biophotonic material over a target skin tissue, wherein the
topical biophotonic material is elastic and comprises at least one
chromophore and a cohesive matrix; and illuminating said biophotonic
material with light having a wavelength that overlaps with an absorption
spectrum of the at least one chromophore; wherein said topical
biophotonic material emits fluorescence at a wavelength and intensity
that treats the acne.

70. (canceled)

71. A method for promoting wound healing comprising: placing a topical
biophotonic material over or within a wound, wherein the topical
biophotonic material is elastic and comprises at least one chromophore
and a cohesive matrix; and illuminating said biophotonic material with
light having a wavelength that overlaps with an absorption spectrum of
the at least one chromophore; wherein said topical biophotonic material
emits fluorescence at a wavelength and intensity that promotes wound
healing.

72. (canceled)

73. A method for promoting skin rejuvenation comprising: placing a
topical biophotonic material over a target skin tissue, wherein the
topical biophotonic material is elastic and comprises at least one
chromophore and a cohesive matrix; and illuminating said biophotonic
material with light having a wavelength that overlaps with an absorption
spectrum of the at least one chromophore; wherein said topical
biophotonic material emits fluorescence at a wavelength and intensity
that promotes skin rejuvenation.

74-79. (canceled)

80. The method of any one of claim 66, 69, 71 or 73, wherein the
biophotonic material is removed after illumination.

81. The method of any one of claim 66, 69, 71 or 73, wherein the
biophotonic material is peelable and is peeled off.

82-85. (canceled)

86. The method of any one of claim 66, 69, 71 or 73, wherein the
chromophore is a xanthene dye.

Description:

[0002] Phototherapy has recently been recognized as having wide range of
applications in both the medical and cosmetic fields including use in
surgery, therapy and diagnostics. For example, phototherapy has been used
to treat cancers and tumors with lessened invasiveness, to disinfect
target sites as an antimicrobial treatment, to promote wound healing, and
for facial skin rejuvenation.

[0003] Photodynamic therapy is a type of phototherapy involving the
application of a photosensitive agent to target tissue then exposing the
target tissue to a light source after a determined period of time during
which the photosensitizer is absorbed by the target tissue. Such
regimens, however, are often associated with undesired side-effects,
including systemic or localized toxicity to the patient or damage to
non-targeted tissue. Moreover, such existing regimens often demonstrate
low therapeutic efficacy due to, for example, the poor selectivity of the
photosensitive agents into the target tissues.

[0004] Therefore, it is an object of the present disclosure to provide new
and improved compositions and methods useful in phototherapy.

[0006] In particular, the biophotonic materials of the present disclosure
include a cohesive matrix, and at least one chromophore, wherein the at
least one chromophore can absorb and emit light from within the
biophotonic material. In certain embodiments of any of the foregoing or
following, the biophotonic material is an elastic material.

[0007] From another aspect, there is provided a topical biophotonic
material comprising: a cohesive matrix, and at least one chromophore
which can absorb and emit light from within the biophotonic material,
wherein the topical biophotonic material is a peelable film.

[0008] From another aspect, there is provided a topical biophotonic
material comprising: a cohesive matrix, and at least one chromophore
which can absorb and emit light from within the biophotonic material,
wherein the topical biophotonic material is elastic.

[0009] From yet another aspect, there is provided a topical biophotonic
material comprising: a cohesive matrix, and at least one chromophore
which can absorb and emit light from within the biophotonic material,
wherein the topical biophotonic material is rigid.

[0010] From another aspect, there is provided a topical biophotonic
material comprising: a cohesive matrix, and at least one chromophore
which can absorb and emit light from within the biophotonic material,
wherein a tear and/or a tensile strength of the topical biophotonic
material is greater than an adhesive strength of the topical biophotonic
material to a surface to which it is applied.

[0011] From a yet further aspect, there is provided a topical biophotonic
material comprising: a cohesive matrix, and at least one chromophore
which can absorb and emit light from within the biophotonic material,
wherein the topical biophotonic material has a well-defined shape under
steady state conditions.

[0012] From another aspect, there is provided a topical biophotonic
material comprising: a cohesive matrix, and at least one chromophore
which can absorb and emit light from within the biophotonic material,
wherein the topical biophotonic material is a mask or a dressing. In
certain embodiments, the mask and/or the dressing has a pre-formed
configuration. In certain embodiments, the mask and/or the dressing is
elastic. In certain embodiments, the mask and/or the dressing is rigid.

[0013] From another aspect, there is provided a biophotonic material
comprising: a cohesive matrix, and at least one chromophore which can
absorb and emit light from within the biophotonic material, wherein the
biophotonic material has a pre-formed configuration which is a shape
and/or a size corresponding with a shape and/or a size of a light source
or lamp to which the biophotonic material can be attached.

[0014] In certain embodiments of the above aspects, the biophotonic
material is a peelable film. In some embodiments, the biophotonic
material is rigid.

[0015] In certain embodiments of any of the foregoing or following, the
biophotonic material has a tear and/or a tensile strength greater than an
adhesive strength of the biophotonic material to a surface to which it is
applied. The adhesive strength may comprise a force required to overcome
static friction.

[0016] In certain embodiments of any of the foregoing or following, the
biophotonic material is at least substantially translucent. The
biophotonic material may be transparent. In some embodiments, the
biophotonic material has a translucency of at least about 40%, about 50%,
about 60%, about 70%, or about 80% in a visible range. Preferably, the
light transmission through the material is measured in the absence of the
at least one chromophore.

[0017] In certain embodiments of any of the foregoing or following, the
biophotonic material has a thickness of about 0.1 mm to about 50 mm,
about 0.5 mm to about 20 mm, or about 1 mm to about 10 mm.

[0018] In certain embodiments of any of the foregoing or following, the
biophotonic material has a pre-formed configuration. In some embodiments,
the pre-formed configuration is a shape and/or a size corresponding with
a shape and/or a size of a body part to which the biophotonic material
can be applied. In some embodiments, the body part to which the material
is applied is a head, scalp, forehead, nose, cheeks, ears, lip, face,
neck, shoulder, arm pit, arm, elbow, hand, finger, abdomen, chest,
stomach, back, sacrum, buttocks, genitals, legs, knee, feet, nails, hair,
toes, or bony prominences, or combinations thereof.

[0019] In certain embodiments of any of the foregoing or following, the
biophotonic material is a mask. In some embodiments, the mask is a face
mask having at least one opening for the eyes, nose or mouth. In certain
embodiments, the mask is disposable. The mask may also be reusable. The
chromophore may at least substantially photobleach after a single use or
single light illumination.

[0020] In certain embodiments of any of the foregoing or following, the
biophotonic material has a pre-formed configuration and the pre-formed
configuration is a shape and/or a size corresponding with a shape and/or
a size of a light source or lamp to which the biophotonic material can be
attached.

[0021] In certain embodiments of any of the foregoing or following, the
biophotonic material can be removed without leaving substantially any
residue on a surface to which the biophotonic material is applied.

[0022] In certain embodiments of any of the foregoing or following, the at
least one chromophore included in the biophotonic material is a
fluorophore. In certain embodiments, the chromophore can absorb and/or
emit light within the visible range. The chromophore may be water
soluble. In certain embodiments, the chromophore can emit light from
around 500 nm to about 700 nm. In some embodiments, the chromophore or
the fluorophore is a xanthene dye. The xanthene dye may be selected from
Eosin Y, Erythrosine B, Fluorescein, Rose Bengal and Phloxin B In some
embodiments, the chromophore is included in the cohesive matrix. In
certain embodiments of any of the foregoing or following, the cohesive
matrix is in particulate form.

[0023] In certain embodiments of any of the foregoing or following, the
cohesive matrix of the biophotonic material comprises at least one
polymer. In some embodiments, the polymer is selected from a cross-linked
polyacrylic polymer, a hyaluronate, a hydrated polymer, a hydrophilic
polymer and a liposoluble polymer. In some embodiments, the cohesive
matrix comprises sodium hyaluronate. In some embodiments, sodium
hyaluronate is present in an amount of about 2% to about 8%.

[0024] In certain embodiments, the cohesive matrix is a liposoluble
polymer, such as silicone. The chromophore(s) may be water soluble and be
within an aqueous phase within the liposoluble polymer. In this case, the
biophotonic material comprises an aqueous phase containing the
chromophore within the liposoluble polymer phase. The aqueous phase may
comprise about 2 wt % to about 40 wt % of the liposoluble polymer phase.
The aqueous phase may be a liquid or a gel. The biophotonic material may
further comprise a stabilizing agent such as CMC or gelatin.

[0025] In certain embodiments, the cohesive matrix comprises gelatin or
chitosan. In certain embodiments, the biophotonic material further
comprises an oxygen-rich compound which may be selected from hydrogen
peroxide, carbamide peroxide and benzoyl peroxide.

[0026] In some embodiments, the chromophore is included in a carrier
medium which can form a cohesive matrix. In some embodiments, the
chromophore can absorb and emit light within the cohesive matrix when
illuminated with light. In some embodiments, the carrier medium is at
least one polymer or a polymer pre-cursor which can form the cohesive
matrix by polymerizing, cross-linking or drying.

[0027] From another aspect, there is provided a topical biophotonic
material comprising a water soluble chromophore within an aqueous
cohesive matrix, and wherein the aqueous cohesive matrix is dispersed
within a liposoluble polymer. In certain embodiments, the liposoluble
polymer is silicone. The aqueous phase may be a liquid or a gel. In
certain embodiments, the aqueous cohesive matrix may be gelatin, water or
carboxymethylcellulose. The chromophore may comprise a fluorophore, such
as a xanthene dye selected from eosin y, fluorescein, erythrosine,
Phloxine b and rose bengal. The aqueous phase may comprise about 2 wt %
to about 40 wt % of the liposoluble polymer phase. In certain
embodiments, the topical biophotonic material may be used to treat
wounds, or to treat or prevent scarring.

[0028] The biophotonic material of any aspects and embodiments of the
disclosure may be used as a mask, dressing or filter. The biophotonic
material of any aspects or embodiments of the disclosure may also be used
for cosmetic or medical treatment of tissue. In some embodiments, the
cosmetic treatment is skin rejuvenation and conditioning, and the medical
treatment is wound healing, periodontal treatment or acne treatment or
treatment of other skin conditions including acne, eczema, psoriasis or
dermatitis. In some aspects, the topical biophotonic material is used for
modulating inflammation, or for promoting angiogenesis.

[0029] The present disclosure also provides containers comprising the
biophotonic material or precursor material according to various
embodiments of the disclosure. In some embodiments, the container
comprises a sealed chamber for holding a biophotonic material, and an
outlet in communication with the chamber for discharging the biophotonic
material from the container, wherein the biophotonic material comprises
at least one chromophore in a carrier medium which can form a cohesive
matrix after being discharged from the sealed chamber. In some
embodiments, the container is a spray can. The container may be opaque.

[0030] The present disclosure also provides kits for preparing or
providing the biophotonic material or precursor according to various
embodiments of the disclosure. In some embodiments, the kit comprises a
first container comprising a first chromophore; and a second component
comprising a thickening agent, wherein the thickening agent can form a
cohesive matrix when mixed with the first component. In some embodiments,
the second container may comprise an oxygen-rich compound.

[0031] The present disclosure also provides methods for biophotonic
treatment comprising applying the topical biophotonic material of the
disclosure to a target tissue and illuminating the material with light.

[0032] From one aspect, there is provided a method for biophotonic
treatment of a skin disorder wherein the method comprises placing a
biophotonic material on or over a target skin tissue, wherein the
biophotonic material is elastic and comprises at least one chromophore
and a cohesive matrix; and illuminating said biophotonic material with
light having a wavelength that overlaps with an absorption spectrum of
the at least one chromophore; wherein said biophotonic material emits
fluorescence at a wavelength and intensity that promotes healing of said
skin disorder. The skin disorder may be selected from acne, eczema,
psoriasis or dermatitis.

[0033] From another aspect, there is provided a method for biophotonic
treatment of a skin disorder comprising: placing a topical biophotonic
material on or over a target skin tissue, wherein the biophotonic
material comprises at least one chromophore and a cohesive matrix, and
wherein a tear and/or tensile strength of the topical biophotonic
material is greater than an adhesive strength of the topical biophotonic
material to a surface to which it is applied; and illuminating said
topical biophotonic material with light having a wavelength that overlaps
with an absorption spectrum of the at least one chromophore; wherein said
biophotonic material emits fluorescence at a wavelength and intensity
that promotes healing of said skin disorder.

[0034] From another aspect, there is provided a method for biophotonic
treatment of acne comprising: placing a topical biophotonic material on
or over a target skin tissue, wherein the topical biophotonic material is
elastic and comprises at least one chromophore and a cohesive matrix; and
illuminating said biophotonic material with light having a wavelength
that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that treats the acne.

[0035] From another aspect, there is provided a method for biophotonic
treatment of acne comprising: placing a topical biophotonic material on
or over a target skin tissue, wherein the topical biophotonic material
comprises at least one chromophore and a cohesive matrix, and wherein a
tear and/or tensile strength of the topical biophotonic material is
greater than an adhesive strength of the topical biophotonic material to
a surface to which it is applied; and illuminating said biophotonic
material with light having a wavelength that overlaps with an absorption
spectrum of the at least one chromophore; wherein said topical
biophotonic material emits fluorescence at a wavelength and intensity
that treats the acne.

[0036] From another aspect, there is provided a method for promoting wound
healing comprising: placing a topical biophotonic material over or within
a wound, wherein the topical biophotonic material is elastic and
comprises at least one chromophore and a cohesive matrix; and
illuminating said biophotonic material with light having a wavelength
that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that promotes wound healing.

[0037] A method for promoting wound healing comprising: placing a topical
biophotonic material over or within a wound, wherein the topical
biophotonic material comprises at least one chromophore and a cohesive
matrix; and wherein a tear and/or tensile strength of the topical
biophotonic material is greater than an adhesive strength of the topical
biophotonic material to a surface to which it is applied; and
illuminating said biophotonic material with light having a wavelength
that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that promotes wound healing.

[0038] From another aspect, there is provided a method for promoting skin
rejuvenation comprising: placing a topical biophotonic material on or
over a target skin tissue, wherein the topical biophotonic material is
elastic and comprises at least one chromophore and a cohesive matrix; and
illuminating said biophotonic material with light having a wavelength
that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that promotes skin rejuvenation.

[0039] From another aspect, there is provided a method for promoting skin
rejuvenation comprising: placing a topical biophotonic material on or
over a target skin tissue, wherein the topical biophotonic material
comprises at least one chromophore and a cohesive matrix; and wherein a
tear and/or tensile strength of the topical biophotonic material is
greater than an adhesive strength of the topical biophotonic material to
a surface to which it is applied; and illuminating said biophotonic
material with light having a wavelength that overlaps with an absorption
spectrum of the at least one chromophore; wherein said topical
biophotonic material emits fluorescence at a wavelength and intensity
that promotes skin rejuvenation.

[0040] In certain embodiments, the biophotonic material is removed after
illumination. In certain embodiments, the biophotonic material is
peelable and is peeled off after illumination. In certain other
embodiments, the biophotonic material is not peelable but can be removed
in one or more pieces. The biophotonic material may be a mask or a
dressing such a face mask or a wound dressing.

[0041] From another aspect, there is provided a method for promoting skin
rejuvenation comprising: placing a topical biophotonic material which is
a mask on or over a target skin tissue, wherein the topical biophotonic
material comprises at least one chromophore and a cohesive matrix; and
illuminating said biophotonic material with light having a wavelength
that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that promotes skin rejuvenation.

[0042] In certain embodiments, the mask is a face mask having at least one
opening for the eyes, nose or mouth. The mask may be disposable or
reusable.

[0043] From another aspect, there is provided a method for promoting wound
healing comprising: placing a topical biophotonic material which is a
dressing over or within a wound, wherein the topical biophotonic material
comprises at least one chromophore and a cohesive matrix; and
illuminating said biophotonic material with light having a wavelength
that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that promotes wound healing.

[0044] From another aspect, there is provided a method for preventing or
treating scarring comprising: placing a topical biophotonic material
which is a membrane over or within a wound, wherein the topical
biophotonic material comprises at least one chromophore and a cohesive
matrix; and illuminating said biophotonic material with light having a
wavelength that overlaps with an absorption spectrum of the at least one
chromophore; wherein said topical biophotonic material emits fluorescence
at a wavelength and intensity that promotes wound healing.

[0045] In certain embodiments, the biophotonic material is left in place
after illumination for re-illumination. In certain embodiments, the
chromophore at least partially photobleaches after illumination. In
certain embodiments, the biophotonic material is illuminated until the
chromophore is at least partially photobleached.

[0046] In certain embodiments, the topical biophotonic material is
illuminated with visible light. In certain embodiments of any of the
foregoing or following, the at least one chromophore included in the
biophotonic material is a fluorophore. In certain embodiments, the
chromophore can absorb and/or emit light within the visible range. The
chromophore may be water soluble. In certain embodiments, the chromophore
can emit light from around 500 nm to about 700 nm. In some embodiments,
the chromophore or the fluorophore is a xanthene dye. The xanthene dye
may be selected from Eosin Y, Erythrosine B, Fluorescein, Rose Bengal and
Phloxin B In some embodiments, the chromophore is included in the
cohesive matrix.

[0047] In certain embodiments of any of the foregoing or following, the
biophotonic material is at least substantially translucent. The
biophotonic material may be transparent. In some embodiments, the
biophotonic material has a translucency of at least about 40%, about 50%,
about 60%, about 70%, or about 80% in a visible range. Preferably, the
light transmission through the material is measured in the absence of the
at least one chromophore. In certain embodiments of any of the foregoing
or following, the biophotonic material has a thickness of about 0.1 mm to
about 50 mm, about 0.5 mm to about 20 mm, or about 1 mm to about 10 mm.

BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Further aspects and advantages of the present invention will become
better understood with reference to the description in association with
the following in which:

[0049] FIG. 1 illustrates the absorption and emission spectra of donor and
acceptor chromophores. The spectral overlap between the absorption
spectrum of the acceptor chromophore and the emission spectrum of the
donor chromophore is also shown.

[0050] FIG. 2 is a schematic of a Jablonski diagram that illustrates the
coupled transitions involved between a donor emission and acceptor
absorbance.

[0051] FIG. 3 is an emission fluorescence spectrum from an activated
biophotonic material according to an embodiment of the present disclosure
(Example 1).

[0053] FIGS. 5a and 5b are emission fluorescence spectra for Eosin Y and
Fluorescein, respectively, and the activating light passing through the
composition, at different concentrations of the chromophores (Example 4).

[0054] FIGS. 6a and 6b are absorbance and emission spectra, respectively,
of Eosin and Fluorescein in a gel (Example 5).

[0055] FIGS. 7a and 7b are absorbance and emission spectra, respectively,
of Eosin, Fluorescein and Rose Bengal in a gel (Example 6).

[0056] FIGS. 8a and 8b are stress-strain curves of cohesive biophotonic
materials according to embodiments of the present disclosure (Example
10).

DETAILED DESCRIPTION

(1) Overview

[0057] The present disclosure provides biophotonic materials and uses
thereof. Biophotonic therapy using these materials would not involve
substantial direct contact of a photosensitive agent (or chromophore)
with the therapeutic target, which includes, but is not limited to, skin,
mucous membranes, wounds, hair and nails. Therefore, undesired side
effects caused by such direct contact may be reduced, minimized, or
prevented. Furthermore, in certain embodiments, phototherapy using the
biophotonic materials of the present disclosure will for instance
rejuvenate the skin by, e.g., promoting collagen synthesis, promote wound
healing, treat skin conditions such as acne, and treat periodontitis.

(2) Definitions

[0058] Before continuing to describe the present disclosure in further
detail, it is to be understood that this disclosure is not limited to
specific compositions or process steps, as such may vary. It must be
noted that, as used in this specification and the appended claims, the
singular form "a", "an" and "the" include plural referents unless the
context clearly dictates otherwise.

[0059] As used herein, the term "about" in the context of a given value or
range refers to a value or range that is within 20%, preferably within
10%, and more preferably within 5% of the given value or range.

[0060] It is convenient to point out here that "and/or" where used herein
is to be taken as specific disclosure of each of the two specified
features or components with or without the other. For example "A and/or
B" is to be taken as specific disclosure of each of (i) A, (ii) B and
(iii) A and B, just as if each is set out individually herein.

[0061] "Biophotonic" means the generation, manipulation, detection and
application of photons in a biologically relevant context. In other
words, biophotonic compositions and materials exert their physiological
effects primarily due to the generation and manipulation of photons.

[0062] "Biophotonic material" is a material which may be activated by
light to produce photons for biologically relevant applications.
Biophotonic materials, as referred to herein, may be cohesive gels,
semi-solids or solids. The biophotonic material can be in the form of,
including, but not limited to, a film or the like, for uses such as a
mask, a dressing or a light attachment. The biophotonic material can be a
composite and include fibres, particulates, ribs, supporting structures,
networks, non-biophotonic layers or biophotonic layers with the same or
different compositions.

[0063] "Cohesive matrix" refers to a medium which is, or which can form, a
self-supporting material e.g. a material with a defined shape under
steady state conditions. This may be due to internal attractive forces.
The property of cohesion in a material can allow the material to be
handled without tearing.

[0065] Terms "chromophore" and "photoactivator" are used herein
interchangeably. A chromophore means a chemical compound, when contacted
by light irradiation, is capable of absorbing the light. The chromophore
readily undergoes photoexcitation and can transfer its energy to other
molecules or emit it as light (fluorescence).

[0066] "Photobleaching" or "photobleaches" means the photochemical
destruction of a chromophore. A chromophore may fully or partially
photobleach.

[0067] The term "actinic light" is intended to mean light energy emitted
from a specific light source (e.g. lamp, LED, or laser) and capable of
being absorbed by matter (e.g. the chromophore or photoactivator). In a
preferred embodiment, the actinic light is visible light.

[0068] A "peel-off" or "peelable" film, membrane or matrix is one that can
be mechanically removed, such as by hand, after application. It can be
removed as a single piece, or as a small number of large pieces.

[0069] "Skin rejuvenation" means a process of reducing, diminishing,
retarding or reversing one or more signs of skin aging or generally
improving the condition of skin. For instance, increasing luminosity of
the skin, reducing pore size, reducing fine lines or wrinkles, improving
thin and transparent skin, improving firmness, improving sagging skin
(such as that produced by bone loss), improving dry skin (which might
itch), reducing or reversing freckles, age spots, spider veins, rough and
leathery skin, fine wrinkles that disappear when stretched, reducing
loose skin, or improving a blotchy complexion. According to the present
disclosure, one or more of the above conditions may be improved or one or
more signs of aging may be reduced, diminished, retarded or even reversed
by certain embodiments of the compositions, methods and uses of the
present disclosure.

[0070] "Wound" means an injury to any tissue, including for example,
acute, subacute, delayed or difficult to heal wounds, and chronic wounds.
Examples of wounds may include both open and closed wounds. Wounds
include, for example, amputations, burns, incisions, excisions, lesions,
lacerations, abrasions, puncture or penetrating wounds, surgical wounds,
amputations, contusions, hematomas, crushing injuries, ulcers (such as
for example pressure, diabetic, venous or arterial), wounds caused by
periodontitis (inflammation of the periodontium).

[0071] Features and advantages of the subject matter hereof will become
more apparent in light of the following detailed description of selected
embodiments, as illustrated in the accompanying figures. As will be
realized, the subject matter disclosed and claimed is capable of
modifications in various respects, all without departing from the scope
of the claims. Accordingly, the drawings and the description are to be
regarded as illustrative in nature, and not as restrictive and the full
scope of the subject matter is set forth in the claims.

(3) Biophotonic Materials

[0072] The present disclosure provides, in a broad sense, topical
biophotonic materials which are cohesive and methods of using the
biophotonic materials. Biophotonic materials can be, in a broad sense,
activated by light (e.g., photons) of specific wavelength. A biophotonic
material according to various embodiments of the present disclosure
contains a cohesive matrix and at least one chromophore in or on the
cohesive matrix which is activated by light and accelerates the
dispersion of light energy, which leads to light carrying on a
therapeutic effect on its own, and/or to the photochemical activation of
other agents contained in the composition (e.g., acceleration in the
breakdown process of peroxide (an oxidant) when such compound is present
in the composition or in contact with the composition, leading to the
formation of oxygen radicals, such as singlet oxygen).

[0073] When a chromophore absorbs a photon of a certain wavelength, it
becomes excited. This is an unstable condition and the molecule tries to
return to the ground state, giving away the excess energy. For some
chromophores, it is favorable to emit the excess energy as light when
returning to the ground state. This process is called fluorescence. The
peak wavelength of the emitted fluorescence is shifted towards longer
wavelengths compared to the absorption wavelengths due to loss of energy
in the conversion process. This is called the Stokes' shift. In the
proper environment (e.g., in a biophotonic material) much of this energy
is transferred to the other components of the biophotonic material or to
the treatment site directly.

[0074] Without being bound to theory, it is thought that fluorescent light
emitted by photoactivated chromophores may have therapeutic properties
due to its femto-, pico-, or nano-second emission properties which may be
recognized by biological cells and tissues, leading to favourable
biomodulation. Furthermore, the emitted fluorescent light has a longer
wavelength and hence a deeper penetration into the tissue than the
activating light. Irradiating tissue with such a broad range of
wavelength, including in some embodiments the activating light which
passes through the composition, may have different and complementary
effects on the cells and tissues. In other words, chromophores are used
in the biophotonic materials of the present disclosure for therapeutic
effect on tissues. This is a distinct application of these photoactive
agents and differs from the use of chromophores as simple stains or as
catalysts for photo-polymerization.

[0075] The biophotonic materials of the present disclosure may have
topical uses such as a mask or a wound dressing, or as an attachment to a
light source, as a waveguide or as a light filter. The cohesive nature of
these biophotonic materials may provide ease of removal from the site of
treatment and hence a faster and less messy treatment. In addition the
biophotonic materials can limit the contact between the chromopore and
the tissue. These materials may be described based on the components
making up the composition. Additionally or alternatively, the
compositions of the present disclosure have functional and structural
properties and these properties may also be used to define and describe
the compositions. Individual components of the biophotonic materials of
the present disclosure, including chromophores, thickening agents and
other optional ingredients, are detailed below.

[0076] The present disclosure also provides a precursor composition to the
material described herein, which will become cohesive on drying, heating,
light exposure, application to tissue or mixing. The precursor
composition comprises at least one chromophore in a carrier medium, or at
least one chromophore and a cohesive matrix.

[0077] (a) Chromophores

[0078] Suitable chromophores can be fluorescent compounds (or stains)
(also known as "fluorochromes" or "fluorophores"). Other dye groups or
dyes (biological and histological dyes, food colorings, carotenoids,
naturally occurring fluorescent and other dyes) can also be used.
Suitable photoactivators can be those that are Generally Regarded As Safe
(GRAS). Advantageously, photoactivators which are not well tolerated by
the skin or other tissues can be included in the biophotonic material of
the present disclosure, as in certain embodiments, the photoactivators
are encapsulated within the cohesive matrix and may not contact the
tissues

[0079] In certain embodiments, the biophotonic material of the present
disclosure comprises a first chromophore which undergoes partial or
complete photobleaching upon application of light. In some embodiments,
the first chromophore absorbs at a wavelength in the range of the visible
spectrum, such as at a wavelength of about 380-800 nm, 380-700, 400-800,
or 380-600 nm. In other embodiments, the first chromophore absorbs at a
wavelength of about 200-800 nm, 200-700 nm, 200-600 nm or 200-500 nm. In
one embodiment, the first chromophore absorbs at a wavelength of about
200-600 nm. In some embodiments, the first chromophore absorbs light at a
wavelength of about 200-300 nm, 250-350 nm, 300-400 nm, 350-450 nm,
400-500 nm, 450-650 nm, 600-700 nm, 650-750 nm or 700-800 nm.

[0080] It will be appreciated to those skilled in the art that optical
properties of a particular chromophore may vary depending on the
chromophore's surrounding medium. Therefore, as used herein, a particular
chromophore's absorption and/or emission wavelength (or spectrum)
corresponds to the wavelengths (or spectrum) measured in a biophotonic
material of the present disclosure.

[0081] The biophotonic material disclosed herein may include at least one
additional chromophore. Combining chromophores may increase
photo-absorption by the combined dye molecules and enhance absorption and
photo-biomodulation selectivity. This creates multiple possibilities of
generating new photosensitive, and/or selective chromophores mixtures.
Thus, in certain embodiments, biophotonic materials of the disclosure
include more than one chromophore. When such multi-chromophore materials
are illuminated with light, energy transfer can occur between the
chromophores. This process, known as resonance energy transfer, is a
widely prevalent photophysical process through which an excited `donor`
chromophore (also referred to herein as first chromophore) transfers its
excitation energy to an `acceptor` chromophore (also referred to herein
as second chromophore). The efficiency and directedness of resonance
energy transfer depends on the spectral features of donor and acceptor
chromophores. In particular, the flow of energy between chromophores is
dependent on a spectral overlap reflecting the relative positioning and
shapes of the absorption and emission spectra. More specifically, for
energy transfer to occur, the emission spectrum of the donor chromophore
must overlap with the absorption spectrum of the acceptor chromophore
(FIG. 1).

[0082] Energy transfer manifests itself through decrease or quenching of
the donor emission and a reduction of excited state lifetime accompanied
also by an increase in acceptor emission intensity. FIG. 2 is a Jablonski
diagram that illustrates the coupled transitions involved between a donor
emission and acceptor absorbance.

[0083] To enhance the energy transfer efficiency, the donor chromophore
should have good abilities to absorb photons and emit photons.
Furthermore, the more overlap there is between the donor chromophore's
emission spectra and the acceptor chromophore's absorption spectra, the
better a donor chromophore can transfer energy to the acceptor
chromophore.

[0084] In certain embodiments, the biophotonic material of the present
disclosure further comprises a second chromophore. In some embodiments,
the first chromophore has an emission spectrum that overlaps at least
about 80%, 50%, 40%, 30%, 20% or 10% with an absorption spectrum of the
second chromophore. In one embodiment, the first chromophore has an
emission spectrum that overlaps at least about 20% with an absorption
spectrum of the second chromophore. In some embodiments, the first
chromophore has an emission spectrum that overlaps at least 1-10%, 5-15%,
10-20%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 50-60%, 55-65% or 60-70%
with an absorption spectrum of the second chromophore.

[0085] % spectral overlap, as used herein, means the % overlap of a donor
chromophore's emission wavelength range with an acceptor chromophore's
absorption wavelength rage, measured at spectral full width quarter
maximum (FWQM). For example, FIG. 1 shows the normalized absorption and
emission spectra of donor and acceptor chromophores. The spectral FWQM of
the acceptor chromophore's absorption spectrum is from about 60 nm (515
nm to about 575 nm). The overlap of the donor chromophore's spectrum with
the absorption spectrum of the acceptor chromophore is about 40 nm (from
515 nm to about 555 nm). Thus, the % overlap can be calculated as 40
nm/60 nm×100=66.6%.

[0086] In some embodiments, the second chromophore absorbs at a wavelength
in the range of the visible spectrum. In certain embodiments, the second
chromophore has an absorption wavelength that is relatively longer than
that of the first chromophore within the range of about 50-250, 25-150 or
10-100 nm.

[0087] The first chromophore can be present in an amount of about
0.001-40% per weight of the biophotonic material. When present, the
second chromophore can be present in an amount of about 0.001-40% per
weight of the biophotonic material. In certain embodiments, the first
chromophore is present in an amount of about 0.001-3%, 0.001-0.01%,
0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%,
12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%,
30-35%, 32.5-37.5%, or 35-40% per weight of the biophotonic material. In
certain embodiments, the second chromophore is present in an amount of
about 0.001-3%, 0.001-0.01%, 0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%,
2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%,
20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40% per
weight of the biophotonic material. In certain embodiments, the total
weight per weight of chromophore or combination of chromophores may be in
the amount of about 0.005-1%, 0.05-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%,
10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%,
27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40.001% per weight of the
biophotonic material.

[0088] The concentration of the chromophore to be used can be selected
based on the desired intensity and duration of the biophotonic activity
from the biophotonic material, and on the desired medical or cosmetic
effect. For example, some dyes such as xanthene dyes reach a `saturation
concentration` after which further increases in concentration do not
provide substantially higher emitted fluorescence. Further increasing the
chromophore concentration above the saturation concentration can reduce
the amount of activating light passing through the matrix. Therefore, if
more fluorescence is required for a certain application than activating
light, a high `saturation` concentration of chromophore can be used.
However, if a balance is required between the emitted fluorescence and
the activating light, a concentration close to or lower than the
saturation concentration can be chosen.

[0089] Suitable chromophores that may be used in the biophotonic materials
of the present disclosure include, but are not limited to the following:

[0099] In certain embodiments, the biophotonic material of the present
disclosure includes any of the chromophores listed above, or a
combination thereof, so as to provide a synergistic biophotonic effect at
the application site.

[0100] Without being bound to any particular theory, a synergistic effect
of the chromophore combinations means that the biophotonic effect is
greater than the sum of their individual effects. Advantageously, this
may translate to increased reactivity of the biophotonic material, faster
or improved treatment time. Also, the treatment conditions need not be
altered to achieve the same or better treatment results, such as time of
exposure to light, power of light source used, and wavelength of light
used. In other words, use of synergistic combinations of chromophores may
allow the same or better treatment without necessitating a longer time of
exposure to a light source, a higher power light source or a light source
with different wavelengths.

[0101] In some embodiments, the material includes Eosin Y as a first
chromophore and any one or more of Rose Bengal, Fluorescein, Erythrosine,
Phloxine B, chlorophyllin as a second chromophore. It is believed that
these combinations have a synergistic effect as they can transfer energy
to one another when activated due in part to overlaps or close proximity
of their absorption and emission spectra. This transferred energy is then
emitted as fluorescence or leads to production of reactive oxygen
species. This absorbed and reemitted light is thought to be transmitted
throughout the composition, and also to be transmitted into the site of
treatment.

[0102] In further embodiments, the material includes the following
synergistic combinations: Eosin Y and Fluorescein; Fluorescein and Rose
Bengal; Erythrosine in combination with Eosin Y, Rose Bengal or
Fluorescein; Phloxine B in combination with one or more of Eosin Y, Rose
Bengal, Fluorescein and Erythrosine. Other synergistic chromophore
combinations are also possible.

[0103] By means of synergistic effects of the chromophore combinations in
the material, chromophores which cannot normally be activated by an
activating light (such as a blue light from an LED), can be activated
through energy transfer from chromophores which are activated by the
activating light. In this way, the different properties of photoactivated
chromophores can be harnessed and tailored according to the cosmetic or
the medical therapy required.

[0104] For example, Rose Bengal can generate a high yield of singlet
oxygen when activated in the presence of molecular oxygen, however it has
a low quantum yield in terms of emitted fluorescent light. Rose Bengal
has a peak absorption around 540 nm and so can be activated by green
light. Eosin Y has a high quantum yield and can be activated by blue
light. By combining Rose Bengal with Eosin Y, one obtains a composition
which can emit therapeutic fluorescent light and generate singlet oxygen
when activated by blue light. In this case, the blue light photoactivates
Eosin Y which transfers some of its energy to Rose Bengal as well as
emitting some energy as fluorescence.

[0105] In some embodiments, the chromophore or chromophores are selected
such that their emitted fluorescent light, on photoactivation, is within
one or more of the green, yellow, orange, red and infrared portions of
the electromagnetic spectrum, for example having a peak wavelength within
the range of about 490 nm to about 800 nm. In certain embodiments, the
emitted fluorescent light has a power density of between 0.005 to about
10 mW/cm2, about 0.5 to about 5 mW/cm2.

[0106] (b) Cohesive Matrix

[0107] The biophotonic materials of the present disclosure comprise a
cohesive matrix made from one or more thickening agents, or a carrier
medium. In other words, the biophotonic material of the present
disclosure comprise one or more thickening agents, or a carrier medium
which can form a cohesive matrix. These agents are present in an amount
and ratio sufficient to provide a desired viscosity, flexibility,
rigidity, tensile strength, tear strength, elasticity, and adhesiveness.
The desired properties may be one of achieving a peelable film, or a
rigid or flexible matrix. The thickening agents are selected so that the
chromophore can remain photoactive in the cohesive matrix. The thickening
agents are also selected according to the optical transparency of the
cohesive matrix which they will form. The cohesive matrix should be able
to transmit sufficient light to activate the at least one chromophore
and, in embodiments where fluorescence is emitted by the activated
chromophore, the cohesive matrix should also be able to transmit the
emitted fluorescent light to tissues. It will be recognized by persons
skilled in the art that the thickening agent is an appropriate medium for
the chromophore selected. For example, the inventors have noted that some
xanthene dyes do not fluoresce in non-hydrated media, so hydrated
polymers or polar solvents may be used. The thickening agents should also
be selected according to the intended use. For example, if the
biophotonic material is to be applied onto tissue, the cohesive matrix is
preferably a biocompatible material, or the cohesive matrix has an
outside layer of a biocompatible material which will interface the
tissue.

[0108] Thickening Agents

[0109] In some embodiments, the content of a thickening agent used to make
the cohesive matrix is from about 0.001% to about 40% (w/w %) of the
total weight. In certain embodiments, the total content of the thickening
agent is about 0.001-0.01%, about 0.005-0.05%, about 0.01-0.1, about
0.05-0.5% about 0.1-1%, about 0.5-5%, about 1-5%, about 2.5-7.5%, about
5-10%, about 7.5-12.5%, about 10-15%, about 12.5-17.5%, or about 15-20%,
or about 15-25%, or about 20-30%, or about 25-35%, or about 30-40%. It
will be recognized by one of skill in the art that the viscosity,
flexibility, rigidity, tensile strength, tear strength, elasticity, and
adhesiveness can be adjusted by varying the content of the thickening
material. Methods of determining viscosity, flexibility, rigidity,
tensile strength, tear strength, elasticity, and adhesiveness are known
in the art.

[0112] Thickening agents can also include carbomers which are a synthetic
high molecular weight polymer of acrylic acid that is crosslinked with
either allylsucrose or allylethers of pentaerythritol having a molecular
weight of about 3×106. The gelation mechanism depends on
neutralization of the carboxylic acid moiety to form a soluble salt. The
polymer is hydrophilic and produces sparkling clear gels when
neutralized. Carbomers are available as fine white powders which disperse
in water to form acidic colloidal suspensions (a 1% dispersion has
approx. pH 3) of low viscosity. Neutralization of these suspensions using
a base, for example sodium, potassium or ammonium hydroxides, low
molecular weight amines and alkanolamines, results in the formation of
clear translucent gels.

[0114] In certain embodiments of the disclosure, the thickening agent or
the carrier medium may comprise gelatin. For example, the cohesive matrix
may comprise at least about 5%, about 5 to about 25 weight %, or about 10
to about 20 weight % gelatin within the cohesive biophotonic material.
Alternatively, a lower weight percentage of gelatin may be used together
with chemical cross-linkers or any other cross-linking means.

[0115] In certain embodiments of the disclosure, the thickening agent or
the carrier medium may comprise sodium hyaluronate, which may be present
in an amount of about 2% to about 14%.

[0116] The biophotonic material of the present disclosure may be water
soluble. Alternatively, the biophotonic material of the present
disclosure may optionally include a water-insoluble or liposoluble
substrate. By "water insoluble", it is meant that the substrate does not
dissolve in or readily break apart upon immersion in water. In some
embodiments, the water-insoluble substrate is the implement or vehicle
for delivering the treatment composition to the skin or target tissue. A
wide variety of materials can be used as the water-insoluble substrate.
The following non-limiting characteristics may be desirable: (i)
sufficient wet strength for use, (ii) sufficient softness, (iii)
sufficient thickness, (iv) appropriate size, (v) air permeability, and
(vi) hydrophilicity.

[0117] Non-limiting examples of suitable water-insoluble substrates which
meet the above criteria include nonwoven substrates, woven substrates,
hydroentangled substrates, air entangled substrates, natural sponges,
synthetic sponges, polymeric netted meshes, and the like. Preferred
embodiments employ nonwoven substrates since they are economical and
readily available in a variety of materials. By "nonwoven", it is meant
that the layer is comprised of fibers which are not woven into a fabric
but rather are formed into a sheet, mat, or pad layer.

[0118] In one embodiment of the disclosure, the thickening agent or the
cohesive agent may comprise a silicone membrane. In this embodiment, the
chromophore or chromophores can be included directly within the silicone
membrane or if they are water soluble within inclusions in the membrane
as an aqueous phase. For example, the aqueous phase may be present as a
micro-emulsion within the silicone. The aqueous phase may be a liquid or
a semi-solid. The aqueous phase may further comprise a stabilizing agent
to stabilize the emulsion such as gelatin or CMC. The aqueous phase may
comprise up to 40 wt % of the silicone polymer phase.

[0121] Hydrogen peroxide (H2O2) is a powerful oxidizing agent,
and breaks down into water and oxygen and does not form any persistent,
toxic residual compound. A suitable range of concentration over which
hydrogen peroxide can be used in the biophotonic material is from about
0.1% to about 3%, about 0.1 to 1.5%, about 0.1% to about 1%, about 1%,
less than about 1%.

[0122] Urea hydrogen peroxide (also known as urea peroxide, carbamide
peroxide or percarbamide) is soluble in water and contains approximately
35% hydrogen peroxide. A suitable range of concentration over which urea
peroxide can be used in the biophotonic material of the present
disclosure is less than about 0.25%, or less than about 0.3%, from 0.001
to 0.25%, or from about 0.3% to about 5%. Urea peroxide breaks down to
urea and hydrogen peroxide in a slow-release fashion that can be
accelerated with heat or photochemical reactions.

[0123] Benzoyl peroxide consists of two benzoyl groups (benzoic acid with
the H of the carboxylic acid removed) joined by a peroxide group. It is
found in treatments for acne, in concentrations varying from 2.5% to 10%.
The released peroxide groups are effective at killing bacteria. Benzoyl
peroxide also promotes skin turnover and clearing of pores, which further
contributes to decreasing bacterial counts and reduce acne. Benzoyl
peroxide breaks down to benzoic acid and oxygen upon contact with skin,
neither of which is toxic. A suitable range of concentration over which
benzoyl peroxide can be used in the matrix biophotonic is from about 2.5%
to about 5%.

[0124] According to certain embodiments, the biophotonic material of the
present disclosure may optionally comprise one or more additional
components, such as oxygen-rich compounds as a source of oxygen radicals.
Peroxide compounds are oxidants that contain the peroxy group
(R--O--O--R), which is a chainlike structure containing two oxygen atoms,
each of which is bonded to the other and a radical or some element. When
a biophotonic material of the present disclosure comprising an oxidant is
illuminated with light, the chromophores are excited to a higher energy
state. When the chromophores' electrons return to a lower energy state,
they emit photons with a lower energy level, thus causing the emission of
light of a longer wavelength (Stokes' shift). In the proper environment,
some of this energy is transferred to oxygen or the reactive hydrogen
peroxide and causes the formation of oxygen radicals, such as singlet
oxygen. The singlet oxygen and other reactive oxygen species generated by
the activation of the biophotonic material are thought to operate in a
hormetic fashion. That is, a health beneficial effect that is brought
about by the low exposure to a normally toxic stimuli (e.g. reactive
oxygen), by stimulating and modulating stress response pathways in cells
of the targeted tissues. Endogenous response to exogenous generated free
radicals (reactive oxygen species) is modulated in increased defense
capacity against the exogenous free radicals and induces acceleration of
healing and regenerative processes. Furthermore, activation of the
oxidant may also produce an antibacterial effect. The extreme sensitivity
of bacteria to exposure to free radicals makes the biophotonic material
of the present disclosure potentially a bactericidal composition.

[0127] Specific bisphenolic antimicrobial agents that can be used in the
disclosure include, but are not limited to: 2,2'-methylene
bis-(4-chlorophenol); 2,4,4'trichloro-2'-hydroxy-diphenyl ether, which is
sold by Ciba Geigy, Florham Park, N.J. under the tradename
Triclosan®; 2,2'-methylene bis-(3,4,6-trichlorophenol);
2,2'-methylene bis-(4-chloro-6-bromophenol);
bis-(2-hydroxy-3,5-dichlorophenyl) sulphide; and
bis-(2-hydroxy-5-chlorobenzyl)sulphide.

[0128] Specific benzoic esters (parabens) that can be used in the
disclosure include, but are not limited to: methylparaben; propylparaben;
butylparaben; ethylparaben; isopropylparaben; isobutylparaben;
benzylparaben; sodium methylparaben; and sodium propylparaben.

[0129] Specific halogenated carbanilides that can be used in the
disclosure include, but are not limited to: 3,4,4'-trichlorocarbanilides,
such as 3-(4-chlorophenyl)-1-(3,4-dichlorphenyl)urea sold under the
tradename Triclocarban® by Ciba-Geigy, Florham Park, N.J.;
3-trifluoromethyl-4,4'-dichlorocarbanilide; and
3,3',4-trichlorocarbanilide.

[0130] Specific polymeric antimicrobial agents that can be used in the
disclosure include, but are not limited to: polyhexamethylene biguanide
hydrochloride; and poly(iminoimidocarbonyl iminoimidocarbonyl
iminohexamethylene hydrochloride), which is sold under the tradename
Vantocil® IB.

[0131] Specific thazolines that can be used in the disclosure include, but
are not limited to that sold under the tradename Micro-Check®; and
2-n-octyl-4-isothiazolin-3-one, which is sold under the tradename
Vinyzene® IT-3000 DIDP.

[0132] Specific trichloromethylthioimides that can be used in the
disclosure include, but are not limited to:
N-(trichloromethylthio)phthalimide, which is sold under the tradename
Fungitrol®; and
N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide, which is sold
under the tradename Vancide®.

[0134] Specific metal salts that can be used in the disclosure include,
but are not limited to, salts of metals in groups 3a-5a, 3b-7b, and 8 of
the periodic table. Specific examples of metal salts include, but are not
limited to, salts of: aluminum; zirconium; zinc; silver; gold; copper;
lanthanum; tin; mercury; bismuth; selenium; strontium; scandium; yttrium;
cerium; praseodymiun; neodymium; promethum; samarium; europium;
gadolinium; terbium; dysprosium; holmium; erbium; thalium; ytterbium;
lutetium; and mixtures thereof. An example of the metal-ion based
antimicrobial agent is sold under the tradename HealthShield®, and is
manufactured by HealthShield Technology, Wakefield, Mass.

[0135] Specific broad-spectrum antimicrobial agents that can be used in
the disclosure include, but are not limited to, those that are recited in
other categories of antimicrobial agents herein.

[0136] Additional antimicrobial agents that can be used in the methods of
the disclosure include, but are not limited to: pyrithiones, and in
particular pyrithione-including zinc complexes such as that sold under
the tradename Octopirox®; dimethyidimethylol hydantoin, which is sold
under the tradename Glydant®;
methylchloroisothiazolinone/methylisothiazolinone, which is sold under
the tradename Kathon CG®; sodium sulfite; sodium bisulfate;
imidazolidinyl urea, which is sold under the tradename Germall 115®;
diazolidinyl urea, which is sold under the tradename Germall 11®;
benzyl alcohol v2-bromo-2-nitropropane-1,3-diol, which is sold under the
tradename Bronopol®; formalin or formaldehyde; iodopropenyl
butylcarbamate, which is sold under the tradename Polyphase P100®;
chloroacetamide; methanamine; methyldibromonitrile glutaronitrile
(1,2-dibromo-2,4-dicyanobutane), which is sold under the tradename
Tektamer®; glutaraldehyde; 5-bromo-5-nitro-1,3-dioxane, which is sold
under the tradename Bronidox®; phenethyl alcohol;
o-phenylphenol/sodium o-phenylphenol sodium hydroxymethylglycinate, which
is sold under the tradename Suttocide A®; polymethoxy bicyclic
oxazolidine; which is sold under the tradename Nuosept C®;
dimethoxane; thimersal; dichlorobenzyl alcohol; captan; chlorphenenesin;
dichlorophene; chlorbutanol; glyceryl laurate; halogenated diphenyl
ethers; 2,4,4'-trichloro-2'-hydroxy-diphenyl ether, which is sold under
the tradename Triclosan® and is available from Ciba-Geigy, Florham
Park, N.J.; and 2,2'-dihydroxy-5,5'-dibromo-diphenyl ether.

[0137] Additional antimicrobial agents that can be used in the methods of
the disclosure include those disclosed by U.S. Pat. Nos. 3,141,321;
4,402,959; 4,430,381; 4,533,435; 4,625,026; 4,736,467; 4,855,139;
5,069,907; 5,091,102; 5,639,464; 5,853,883; 5,854,147; 5,894,042; and
5,919,554, and U.S. Pat. Appl. Publ. Nos. 20040009227 and 20110081530.

[0138] (4) Optical Properties of the Biophotonic Materials

[0139] In certain embodiments, biophotonic materials of the present
disclosure are substantially transparent or translucent. The %
transmittance of the biophotonic material can be measured in the range of
wavelengths from 250 nm to 800 nm using, for example, a Perkin-Elmer
Lambda 9500 series UV-visible spectrophotometer. In some embodiments,
transmittance within the visible range is measured and averaged. In some
other embodiments, transmittance of the biophotonic material is measured
with the chromophore omitted. As transmittance is dependent upon
thickness, the thickness of each sample can be measured with calipers
prior to loading in the spectrophotometer. Transmittance values can be
normalized according to

where tactual specimen thickness, t2=thickness to which
transmittance measurements can be normalized. In the art, transmittance
measurements are usually normalized to 1 cm.

[0140] In certain embodiments, the biophotonic materials are substantially
opaque. In these embodiments, the biophotonic materials may include light
transmitting structures such as fibres, particles, networks, which are
made of materials which can transmit light. The light transmitting
structures can be waveguides such as optical fibres.

[0141] In some embodiments, the biophotonic material has a transmittance
that is more than about 20%, 30%, 40%, 50%, 60%, 70%, or 75% within the
visible range. In some embodiments, the transmittance exceeds 40%, 41%,
42%, 43%, 44%, or 45% within the visible range.

[0142] (5) Forms of the Biophotonic Materials

[0143] The biophotonic materials of the present disclosure may be in the
form of a cohesive film or matrix containing at least one chromophore.
The cohesive film or matrix may be a cohesive gel, or a paste, a putty, a
semi-solid, or a solid.

[0144] The biophotonic materials of the present disclosure may be
deformable. They may be elastic or non-elastic (i.e. flexible or rigid).
The biophotonic materials, for example, may be in a peel-off form
(`peelable`) to provide ease and speed of use. In certain embodiments,
the tear strength and/or tensile strength of the peel-off form is greater
than its adhesion strength. This may help handleability of the material.
It will be recognized by one of skill in the art that the properties of
the peel-off biophotonic material such as cohesiveness, flexibility,
elasticity, tensile strength, and tearing strength, can be determined
and/or adjusted by methods known in the art such as by selecting suitable
thickening agents and adapting their relative ratios.

[0145] The biophotonic material may be in a pre-formed shape. In certain
embodiments, the pre-formed shape is in the form of, including, but not
limited to, a film, a face mask, a patch, a dressing, or bandage. In
certain embodiments, the pre-formed shapes can be customized for the
individual user by trimming to size. In certain embodiments, perforations
are provided around the perimeter of the pre-formed shape to facilitate
trimming. In certain embodiments, the pre-shaping can be performed
manually or by mechanical means such as 3-D printing. In the case of the
3-D printing the size of the area to be treated can be imaged, such as a
wound or a face, then a 3-D printer configured to build or form a
cohesive biophotonic material to match the size and shape of the imaged
treatment area.

[0146] A biophotonic material of the disclosure can be configured with a
shape and/or size for application to a desired portion of a subject's
body. For example, the biophotonic material can be shaped and sized to
correspond with a desired portion of the body to receive the biophotonic
treatment. Such a desired portion of skin can be selected from, but not
limited to, the group consisting of a skin, head, forehead, scalp, nose,
cheeks, lips, ears, face, neck, shoulder, arm pit, arm, elbow, hand,
finger, abdomen, chest, stomach, back, buttocks, sacrum, genitals, legs,
knee, feet, toes, nails, hair, any boney prominences, and combinations
thereof, and the like. Thus, the biophotonic material of the disclosure
can be shaped and sized to be applied to any portion of skin on a
subject's body. For example, the biophotonic material can be sock, hat,
glove or mitten shaped. In embodiments where the biophotonic material is
elastic or rigid, it can be peeled-off without leaving any residue on the
tissue.

[0147] In certain embodiments, the biophotonic material is in the form of
an elastic and peelable face mask, which may be pre-formed. In other
embodiments, the biophotonic material is in the form of a non-elastic
(rigid) face mask, which may also be pre-formed. The mask can have
openings for one or more of the eyes, nose and mouth. In a further
embodiment, the openings are protected with a covering, or the exposed
skin such as on the nose, lips or eyes are protected using for example
cocoa butter. In certain embodiments, the pre-formed face mask is
provided in the form of multiple parts, e.g., an upper face part and a
lower face part. In certain embodiments, the uneven proximity of the face
to a light source is compensated for, e.g., by adjusting the thickness of
the mask, or by adjusting the amount of chromophore in the different
areas of the mask, or by blocking the skin in closest proximity to the
light. In certain embodiments, the pre-formed shapes come in a one-size
fits all form.

[0148] In certain aspects, the mask (or patch) is not pre-formed and is
applied e.g., by spreading a composition making up the mask (or patch),
on the skin or target tissue, or alternatively by spraying, smearing,
dabbing or rolling the composition on target tissue. It can then be
converted to a peel-off form after application, by means such as, but not
limited to, drying, illumination with light, change in temperature or pH
upon application to the skin or tissue. The mask (or patch) can then be
peeled off without leaving any flakes on the skin or tissue, preferably
without wiping or washing.

[0149] In certain aspects, the biophotonic material may have shape memory
properties. For example, the biophotonic material can include a shape
memory material, such as a shape memory polymer whose original shape is
reverted to on activation by light. The original shape can be a flat or
concave configuration which allows the film/matrix to be readily peeled
off the tissue. The shape memory material may be included as a layer
attached to the biophotonic material, or integrated with the biophotonic
material.

[0150] In certain aspects, the biophotonic material forms part of a
composite and can include fibres, particulates, non-biophotonic layers or
biophotonic layers with the same or different compositions.

[0151] In certain embodiments, the biophotonic material may comprise a
plurality of waveguides extending at least partially through the
biophotonic material or contained at least partially within the
biophotonic material. The waveguides can be attached to a light source to
thereby illuminate the biophotonic material from within. The biophotonic
material may further include the light source attached to the waveguides.
The waveguides can be optical fibres which can transmit light, not only
from their ends, but also from their body. For example, made of
polycarbonate or polymethylmethacrylate or any other suitable material.

[0152] In a different embodiment, the biophotonic material comprises a
layer of a woven or non-woven fabric dressing or a mask. Waveguides or a
light source may be included within the dressing or mask fabric. For
example, the dressing or mask fabric can be in the form of an envelope
which receives the biophotonic material, and which comprises at least one
light emitting surface.

[0153] In certain aspects, the biophotonic material is formed as a filter.
For example, the biophotonic material can be made to have a shape and a
size which can be connected to, or spaced from, a light emitting surface
of a lamp. In one embodiment, the lamp can be a hand-held lamp such as a
torch or a dentist's curing lamp. The lamp with the biophotonic filter
can then be used to treat tissue sites of patient in a contacting or
non-contacting manner. In this embodiment, the filter has a body having a
first end which is sized and shaped to be connectable to a light emitting
surface, and a second end shaped to treat tissues.

[0154] In certain aspects, the biophotonic material is formed as a
waveguide. In certain embodiments, at least one chromophore is included
in an elongate solid matrix having good light propagation properties and
appropriate mechanical properties. The waveguide may be flexible. The
waveguide can be shaped as an optical fibre. Such an optical fibre can be
connected to a light source, and the at least one chromophore in the
cohesive matrix activated by the light source to deliver therapeutic
fluorescent light to hard to reach places, such as internal cavities and
periodontal pockets. Polymethylmethacrylate is an example of an
appropriate cohesive matrix for use as a biophotonic waveguide. Such a
waveguide may additionally include a coating to prevent light dissipation
from along its length.

[0155] In other aspects, the biophotonic material comprising at least one
chromophore and a cohesive matrix is in the form of particulates.
Material processing techniques known in the art can be used to form
particulates of any shape or size. These particulates can be contained in
semi-solid or liquid preparations. For example, such biophotonic
particulates can be used in skin preparations such as creams, emulsions
to provide therapeutic effect to the skin. In this case, a biocompatible
solid matrix is used and can be used to encapsulate all types of
chromophores, even those not well tolerated by the skin.

[0156] The biophotonic materials of the present disclosure may have a
thickness of from about 0.1 mm to about 50 mm, about 0.5 mm to about 20
mm, or about 1 mm to about 10 mm. It will be appreciated that the
thickness of the biophotonic materials will vary based on the intended
use. In some embodiments, the biophotonic material has a thickness of
from about 0.1-1 mm. In some embodiments, the biophotonic material has a
thickness of about 0.5-1.5 mm, about 1-2 mm, about 1.5-2.5 mm, about 2-3
mm, about 2.5-3.5 mm, about 3-4 mm, about 3.5-4.5 mm, about 4-5 mm, about
4.5-5.5 mm, about 5-6 mm, about 5.5-6.5 mm, about 6-7 mm, about 6.5-7.5
mm, about 7-8 mm, about 7.5-8.5 mm, about 8-9 mm, about 8.5-9.5, about
9-10 mm, about 10-11 mm, about 11-12 mm, about 12-13 mm, about 13-14 mm,
about 14-15 mm, about 15-16 mm, about 16-17 mm, about 17-18 mm, about
18-19 mm, about 19-20 mm, about 20-22 mm, about 22-24 mm, about 24-26 mm,
about 26-28 mm, about 28-30 mm, about 30-35 mm, about 35-40 mm, about
40-45 mm, about 45-50 mm.

[0157] The tensile strength of the biophotonic materials will vary based
on the intended use. The tensile strength can be determined by performing
a tensile test and recording the force and displacement. These are then
converted to stress (using cross sectional area) and strain; the highest
point of the stress-strain curve is the "ultimate tensile strength." In
some embodiments, tensile strength can be characterized using a 500N
capacity tabletop mechanical testing system (#5942R4910, Instron®)
with a 5N maximum static load cell (#102608, Instron). Pneumatic side
action grips can be used to secure the samples (#2712-019, Instron). In
some embodiments, a constant extension rate (for example, of about 2
mm/min) until failure can be applied and the tensile strength is
calculated from the stress vs. strain data plots. In some embodiments,
the tensile strength can be measured using methods as described in or
equivalent to those described in American Society for Testing and
Materials tensile testing methods such as ASTM D638, ASTM D882 and ASTM
D412.

[0158] In some embodiments, the biophotonic material has a tensile
strength of from about 1-50 kPa, 1 to about 1000 kPa, 1 to about 500 kPa,
50 kPa to about 600 kPa. In some embodiments, the tensile strength is
from about 75 kPa to about 500 kPa, from about 100 kPa to about 200 kPa,
100-300 kPa, 400 kPa, from about 150 kPa to about 350 kPa, or from about
200 kPa to about 300 kPa.

[0159] In some embodiments, the tensile strength is at least about 50 kPa,
at least about 75 kPa, at least about 100 kPa, at least about 150 kPa, at
least about 200 kPa, at least about 250 kPa, at least about 300 kPa, at
least about 350 kPa, at least about 400 kPa, at least about 450 kPa, at
least about 500 kPa, at least about 550 kPa or at least about 600 kPa.

[0160] In some embodiments, the tensile strength of the biophotonic
material is up to about 8 MPa.

[0161] The tear strength of the biophotonic material will vary depending
on the intended use. The tear strength property of the biophotonic
material can be tested using a 500N capacity tabletop mechanical testing
system (#5942R4910, Instron) with a 5N maximum static load cell (#102608,
Instron). Pneumatic side action grips can be used to secure the samples
(#2712-019, Instron). Samples can be tested with a constant extension
rate (for example, of about 2 mm/min) until failure. In accordance with
the invention, tear strength is calculated as the force at failure
divided by the average thickness (N/mm).

[0162] In some embodiments, the biophotonic material has a tear strength
of from about 0.1 N/mm to about 1 N/mm. In some embodiments, the tear
strength is from about 0.20 N/mm to about 0.40 N/mm, from about 0.25 N/mm
to about 0.35 N/mm, from about 0.25 N/mm to about 0.45 N/mm, from about
0.35 N/mm to about 0.535 N/mm, from about 0.45 N/mm to about 0.65 N/mm,
from about 0.55 N/mm to about 0.75 N/mm, from about 0.65 N/mm to about
0.85 N/mm, from about 0.75 N/mm to about 0.95 N/mm.

[0163] In some embodiments, the tear strength is at least about 0.10 N/mm,
at least about 0.15 N/mm, at least about 0.20 N/mm, at least about 0.25
N/mm, at least about 0.30 N/mm, at least about 0.35 N/mm, at least about
0.40 N/mm, at least about 0.45 N/mm, at least about 0.55 N/mm or at least
about 1 N/mm.

[0164] The adhesion strength of the biophotonic material will vary
depending on the intended use. Adhesion strength can be determined in
accordance with ASTM D-3330-78, PSTC-101 and is a measure of the force
required to remove a biophotonic material from a test panel at a specific
angle and rate of removal. In some embodiments, a predetermined size of a
biophotonic material is applied to a horizontal surface of a clean glass
test plate. A hard rubber roller is used to firmly apply the piece and
remove all discontinuities and entrapped air. The free end of the piece
of biophotonic material is then doubled back nearly touching itself so
that the angle of removal of the piece from the glass plate will be 180
degrees. The free end of the piece of biophotonic material is attached to
the adhesion tester scale (e.g. an Instron tensile tester or Harvey
tensile tester). The test plate is then clamped in the jaws of the
tensile testing machine capable of moving the plate away from the scale
at a predetermined constant rate. The scale reading in kg is recorded as
the biophotonic material is peeled from the glass surface.

[0165] In some embodiments, the adhesion strength can be measured by
taking into account the static friction of the biophotonic material. For
some embodiments of the cohesive biophotonic materials of the present
disclosure, the adhesive properties are linked to their levels of static
friction, or stiction. In these cases, the adhesion strength can be
measured by placing the sample on a test surface and pulling one end of
the sample at an angle of approximately 0° (substantially parallel
to the surface) whilst applying a known downward force (e.g. a weight) on
the sample and measuring the weight at which the sample slips from the
surface. The normal force Fn, is the force exerted by each surface
on the other in a perpendicular (normal) direction to the surface and is
calculated by multiplying the combined weight of the sample and the
weight by the gravity constant (g) (9.8 m/s2). The biophotonic
material with the weight on top is then pulled away from a balance until
the biophotonic material slips from the surface and the weight is
recorded on the scale. The weight recorded on the scale is equivalent to
the force required to overcome the friction. The force of friction
(Ff) is then calculated by multiplying the weight recorded on the
scale by g. Since Fr≦μFn (Coulomb's friction law),
the friction coefficient μ can be obtained by dividing
Fr/Fn. The stress required to shear a material from a surface
(adhesion strength) can then be calculated from the friction coefficient,
by multiplying the weight of the material by the friction coefficient.

[0166] In some embodiments, the biophotonic material has an adhesion
strength that is less than its tensile strength. In some embodiments, the
biophotonic material has an adhesion strength that is less than its tear
strength.

[0167] In some embodiments, the biophotonic material has an adhesion
strength of from about 0.01 N/mm to about 0.60 N/mm. In some embodiments,
the adhesion strength is from about 0.20 N/mm to about 0.40 N/mm, or from
about 0.25 N/mm to about 0.35 N/mm. In some embodiments, the adhesion
strength is less than about 0.10 N/mm, less than about 0.15 N/mm, less
than about 0.20 N/mm, less than about 0.25 N/mm, less than about 0.30
N/mm, less than about 0.35 N/mm, less than about 0.40 N/mm, less than
about 0.45 N/mm, less than about 0.55 N/mm or less than about 0.60 N/mm.

[0168] (6) Methods of Use

[0169] The biophotonic materials of the present disclosure may have
cosmetic and/or medical benefits. They can be used to promote skin
rejuvenation and skin conditioning, promote the treatment of a skin
disorder such as acne, eczema or psoriasis, promote tissue repair, and
promote wound healing including periodontitis pockets. They can be used
to treat acute inflammation. Acute inflammation can present itself as
pain, heat, redness, swelling and loss of function. It includes those
seen in allergic reactions such as insect bites e.g.; mosquito, bees,
wasps, poison ivy, or post-ablative treatment.

[0171] In certain embodiments, the present disclosure provides a method
for providing skin rejuvenation or for improving skin condition, treating
a skin disorder, preventing or treating scarring, and/or accelerating
wound healing and/or tissue repair, the method comprising: applying a
biophotonic material of the present disclosure to the area of the skin or
tissue in need of treatment, and illuminating the biophotonic material
with light having a wavelength that overlaps with an absorption spectrum
of the chromophore(s) present in the biophotonic material.

[0172] In the methods of the present disclosure, any source of actinic
light can be used. Any type of halogen, LED or plasma arc lamp, or laser
may be suitable. The primary characteristic of suitable sources of
actinic light will be that they emit light in a wavelength (or
wavelengths) appropriate for activating the one or more photoactivators
present in the composition. In one embodiment, an argon laser is used. In
another embodiment, a potassium-titanyl phosphate (KTP) laser (e.g. a
GreenLight® laser) is used. In yet another embodiment, a LED lamp such
as a photocuring device is the source of the actinic light. In yet
another embodiment, the source of the actinic light is a source of light
having a wavelength between about 200 to 800 nm. In another embodiment,
the source of the actinic light is a source of visible light having a
wavelength between about 400 and 600 nm. In another embodiment, the
source of the actinic light is a source of visible light having a
wavelength between about 400 and 700 nm. In yet another embodiment, the
source of the actinic light is blue light. In yet another embodiment, the
source of the actinic light is red light. In yet another embodiment, the
source of the actinic light is green light. Furthermore, the source of
actinic light should have a suitable power density. Suitable power
density for non-collimated light sources (LED, halogen or plasma lamps)
are in the range from about 0.1 mW/cm2 to about 200 mW/cm2.
Suitable power density for laser light sources are in the range from
about 0.5 mW/cm2 to about 0.8 mW/cm2.

[0173] In some embodiments of the methods of the present disclosure, the
light has an energy at the subject's skin surface of between about 0.1
mW/cm2 and about 500 mW/cm2, or 0.1-300 mW/cm2, or 0.1-200
mW/cm2, wherein the energy applied depends at least on the condition
being treated, the wavelength of the light, the distance of the skin from
the light source and the thickness of the biophotonic material. In
certain embodiments, the light at the subject's skin is between about
1-40 mW/cm2, or 20-60 mW/cm2, or 40-80 mW/cm2, or 60-100
mW/cm2, or 80-120 mW/cm2, or 100-140 mW/cm2, or 30-180
mW/cm2, or 120-160 mW/cm2, or 140-180 mW/cm2, or 160-200
mW/cm2, or 110-240 mW/cm2, or 110-150 mW/cm2, or 190-240
mW/cm2.

[0174] The activation of the chromophore(s) within the biophotonic
material may take place almost immediately on illumination (femto- or
pico seconds). A prolonged exposure period may be beneficial to exploit
the synergistic effects of the absorbed, reflected and reemitted light of
the biophotonic material of the present disclosure and its interaction
with the tissue being treated. In one embodiment, the time of exposure to
actinic light of the tissue or skin or biophotonic material is a period
between 1 minute and 5 minutes. In another embodiment, the time of
exposure to actinic light of the tissue or skin or biophotonic material
is a period between 1 minute and 5 minutes. In some other embodiments,
the biophotonic material is illuminated for a period between 1 minute and
3 minutes. In certain embodiments, light is applied for a period of 1-30
seconds, 15-45 seconds, 30-60 seconds, 0.75-1.5 minutes, 1-2 minutes,
1.5-2.5 minutes, 2-3 minutes, 2.5-3.5 minutes, 3-4 minutes, 3.5-4.5
minutes, 4-5 minutes, 5-10 minutes, 10-15 minutes, 15-20 minutes, or
20-30 minutes. The treatment time may range up to about 90 minutes, about
80 minutes, about 70 minutes, about 60 minutes, about 50 minutes, about
40 minutes or about 30 minutes. It will be appreciated that the treatment
time can be adjusted in order to maintain a dosage by adjusting the rate
of fluence delivered to a treatment area. For example, the delivered
fluence may be about 4 to about 60 J/cm2, about 10 to about 60
J/cm2, about 10 to about 50 J/cm2, about 10 to about 40
J/cm2, about 10 to about 30 J/cm2, about 20 to about 40
J/cm2, about 15 J/cm2 to 25 J/cm2, or about 10 to about 20
J/cm2.

[0175] In certain embodiments, the biophotonic material may be
re-illuminated at certain intervals. In yet another embodiment, the
source of actinic light is in continuous motion over the treated area for
the appropriate time of exposure. In yet another embodiment, the
biophotonic composition may be illuminated until the biophotonic
composition is at least partially photobleached or fully photobleached.

[0176] In certain embodiments, the chromophore(s) in the cohesive matrix
can be photoexcited by ambient light including from the sun and overhead
lighting. In certain embodiments, the chromophore(s) can be
photoactivated by light in the visible range of the electromagnetic
spectrum. The light can be emitted by any light source such as sunlight,
light bulb, an LED device, electronic display screens such as on a
television, computer, telephone, mobile device, flashlights on mobile
devices. In the methods of the present disclosure, any source of light
can be used. For example, a combination of ambient light and direct
sunlight or direct artificial light may be used. Ambient light can
include overhead lighting such as LED bulbs, fluorescent bulbs etc, and
indirect sunlight.

[0177] In the methods of the present disclosure, the biophotonic material
may be removed from the skin following application of light. In some
embodiments the biophotonic material is peeled off from the skin
following application of light. In some embodiments, the biophotonic
material is removed as a single piece from the skin following application
of light. In other embodiments, the biophotonic material is left on the
tissue for an extended period of time and re-activated with direct or
ambient light at appropriate times to treat the condition.

[0178] In certain embodiments of the method of the present disclosure, the
biophotonic material can be applied to the tissue, such as on the face,
once, twice, three times, four times, five times or six times a week,
daily, or at any other frequency. The total treatment time can be one
week, two weeks, three weeks, four weeks, five weeks, six weeks, seven
weeks, eight weeks, nine weeks, ten weeks, eleven weeks, twelve weeks, or
any other length of time deemed appropriate. In certain embodiments, the
total tissue area to be treated may be split into separate areas (cheeks,
forehead), and each area treated separately. For example, the composition
may be applied topically to a first portion, and that portion illuminated
with light, and the biophotonic composition then removed. Then the
composition is applied to a second portion, illuminated and removed.
Finally, the composition is applied to a third portion, illuminated and
removed.

[0179] In certain embodiments, the biophotonic material can be used
following wound closure to optimize scar revision. In this case, the
biophotonic material may be applied at regular intervals such as once a
week, or at an interval deemed appropriate by the physician.

[0180] In certain embodiments, the biophotonic material can be used
following acne treatment to maintain the condition of the treated skin.
In this case, the biophotonic material may be applied at regular
intervals such as once a week, or at an interval deemed appropriate by
the physician.

[0181] In certain embodiments, the biophotonic material can be used
following ablative skin rejuvenation treatment to maintain the condition
of the treated skin. In this case, the biophotonic material may be
applied at regular intervals such as once a week, or at an interval
deemed appropriate by the physician.

[0182] In the methods of the present disclosure, additional components may
optionally be included in the biophotonic materials or used in
combination with the biophotonic materials. Such additional components
include, but are not limited to, healing factors, antimicrobials,
oxygen-rich agents, wrinkle fillers such as botox, hyaluronic acid and
polylactic acid, fungal, anti-bacterial, anti-viral agents and/or agents
that promote collagen synthesis. These additional components may be
applied to the skin in a topical fashion, prior to, at the same time of;
and/or after topical application of the biophotonic materials of the
present disclosure. Suitable healing factors comprise compounds that
promote or enhance the healing or regenerative process of the tissues on
the application site. During the photoactivation of a biophotonic
material of the present disclosure, there may be an increase of the
absorption of molecules of such additional components at the treatment
site by the skin or the mucosa. In certain embodiments, an augmentation
in the blood flow at the site of treatment can observed for a period of
time. An increase in the lymphatic drainage and a possible change in the
osmotic equilibrium due to the dynamic interaction of the free radical
cascades can be enhanced or even fortified with the inclusion of healing
factors. Healing factors may also modulate the biophotonic output from
the biophotonic composition such as photobleaching time and profile, or
modulate leaching of certain ingredients within the composition. Suitable
healing factors include, but are not limited to glucosamines, allantoins,
saffron, agents that promote collagen synthesis, anti-fungal,
anti-bacterial, anti-viral agents and wound healing factors such as
growth factors.

[0183] (i) Skin Rejuvenation

[0184] The biophotonic material of the present disclosure may be useful in
promoting skin rejuvenation or improving skin condition and appearance.
The dermis is the second layer of skin, containing the structural
elements of the skin, the connective tissue. There are various types of
connective tissue with different functions. Elastin fibers give the skin
its elasticity, and collagen gives the skin its strength.

[0185] The junction between the dermis and the epidermis is an important
structure. The dermal-epidermal junction interlocks forming finger-like
epidermal ridges. The cells of the epidermis receive their nutrients from
the blood vessels in the dermis. The epidermal ridges increase the
surface area of the epidermis that is exposed to these blood vessels and
the needed nutrients.

[0186] The aging of skin comes with significant physiological changes to
the skin. The generation of new skin cells slows down, and the epidermal
ridges of the dermal-epidermal junction flatten out. While the number of
elastin fibers increases, their structure and coherence decreases. Also
the amount of collagen and the thickness of the dermis decrease with the
ageing of the skin.

[0187] Collagen is a major component of the skin's extracellular matrix,
providing a structural framework. During the aging process, the decrease
of collagen synthesis and insolubilization of collagen fibers contribute
to a thinning of the dermis and loss of the skin's biomechanical
properties.

[0188] The physiological changes to the skin result in noticeable aging
symptoms often referred to as chronological-, intrinsic- and
photo-ageing. The skin becomes drier, roughness and scaling increase, the
appearance becomes duller, and most obviously fine lines and wrinkles
appear. Other symptoms or signs of skin aging include, but are not
limited to, thinning and transparent skin, loss of underlying fat
(leading to hollowed cheeks and eye sockets as well as noticeable loss of
firmness on the hands and neck), bone loss (such that bones shrink away
from the skin due to bone loss, which causes sagging skin), dry skin
(which might itch), inability to sweat sufficiently to cool the skin,
unwanted facial hair, freckles, age spots, spider veins, rough and
leathery skin, fine wrinkles that disappear when stretched, loose skin, a
blotchy complexion.

[0189] The dermal-epidermal junction is a basement membrane that separates
the keratinocytes in the epidermis from the extracellular matrix, which
lies below in the dermis. This membrane consists of two layers: the basal
lamina in contact with the keratinocytes, and the underlying reticular
lamina in contact with the extracellular matrix. The basal lamina is rich
in collagen type IV and laminin, molecules that play a role in providing
a structural network and bioadhesive properties for cell attachment.

[0190] Laminin is a glycoprotein that only exists in basement membranes.
It is composed of three polypeptide chains (alpha, beta and gamma)
arranged in the shape of an asymmetric cross and held together by
disulfide bonds. The three chains exist as different subtypes which
result in twelve different isoforms for laminin, including Laminin-1 and
Laminin-5.

[0191] The dermis is anchored to hemidesmosomes, specific junction points
located on the keratinocytes, which consist of a-integrins and other
proteins, at the basal membrane keratinocytes by type VII collagen
fibrils. Laminins, and particularly Laminin-5, constitute the real anchor
point between hemidesmosomal transmembrane proteins in basal
keratinocytes and type VII collagen.

[0192] Laminin-5 synthesis and type VII collagen expression have been
proven to decrease in aged skin. This causes a loss of contact between
dermis and epidermis, and results in the skin losing elasticity and
becoming saggy.

[0193] Recently another type of wrinkles, generally referred to as
expression wrinkles, got general recognition. These wrinkles require loss
of resilience, particularly in the dermis, because of which the skin is
no longer able to resume its original state when facial muscles which
produce facial expressions exert stress on the skin, resulting in
expression wrinkles.

[0194] The biophotonic material of the present disclosure and methods of
the present disclosure promote skin rejuvenation. In certain embodiments,
the biophotonic material and methods of the present disclosure promote
skin condition such as skin luminosity, reduction of pore size, reducing
blotchiness, making even skin tone, reducing dryness, and tightening of
the skin. In certain embodiments, the biophotonic material and methods of
the present disclosure promote collagen synthesis. In certain other
embodiments, the biophotonic material and methods of the present
disclosure may reduce, diminish, retard or even reverse one or more signs
of skin aging including, but not limited to, appearance of fine lines or
wrinkles, thin and transparent skin, loss of underlying fat (leading to
hollowed cheeks and eye sockets as well as noticeable loss of firmness on
the hands and neck), bone loss (such that bones shrink away from the skin
due to bone loss, which causes sagging skin), dry skin (which might
itch), inability to sweat sufficiently to cool the skin, unwanted facial
hair, freckles, age spots, spider veins, rough and leathery skin, fine
wrinkles that disappear when stretched, loose skin, or a blotchy
complexion. In certain embodiments, the biophotonic material and methods
of the present disclosure may induce a reduction in pore size, enhance
sculpturing of skin subsections, and/or enhance skin translucence.

[0196] For instance, it was discovered that intake of vitamin C, iron, and
collagen can effectively increase the amount of collagen in skin or bone.
See, e.g., U.S. Patent Application Publication 20090069217. Examples of
the vitamin C include an ascorbic acid derivative such as L-ascorbic acid
or sodium L-ascorbate, an ascorbic acid preparation obtained by coating
ascorbic acid with an emulsifier or the like, and a mixture containing
two or more of those vitamin Cs at an arbitrary rate. In addition,
natural products containing vitamin C such as acerola and lemon may also
be used. Examples of the iron preparation include: an inorganic iron such
as ferrous sulfate, sodium ferrous citrate, or ferric pyrophosphate; an
organic iron such as heme iron, ferritin iron, or lactoferrin iron; and a
mixture containing two or more of those irons at an arbitrary rate. In
addition, natural products containing iron such as spinach or liver may
also be used. Moreover, examples of the collagen include: an extract
obtained by treating bone, skin, or the like of a mammal such as bovine
or swine with an acid or alkaline; a peptide obtained by hydrolyzing the
extract with a protease such as pepsin, trypsin, or chymotrypsin; and a
mixture containing two or more of those collagens at an arbitrary rate.
Collagens extracted from plant sources may also be used.

[0201] The biophotonic materials and methods of the present disclosure may
be used to treat acne. As used herein, "acne" means a disorder of the
skin caused by inflammation of skin glands or hair follicles. The
biophotonic materials and methods of the disclosure can be used to treat
acne at early pre-emergent stages or later stages where lesions from acne
are visible. Mild, moderate and severe acne can be treated with
embodiments of the biophotonic compositions and methods. Early
pre-emergent stages of acne usually begin with an excessive secretion of
sebum or dermal oil from the sebaceous glands located in the
pilosebaceous apparatus. Sebum reaches the skin surface through the duct
of the hair follicle. The presence of excessive amounts of sebum in the
duct and on the skin tends to obstruct or stagnate the normal flow of
sebum from the follicular duct, thus producing a thickening and
solidification of the sebum to create a solid plug known as a comedone.
In the normal sequence of developing acne, hyperkeratinazation of the
follicular opening is stimulated, thus completing blocking of the duct.
The usual results are papules, pustules, or cysts, often contaminated
with bacteria, which cause secondary infections. Acne is characterized
particularly by the presence of comedones, inflammatory papules, or
cysts. The appearance of acne may range from slight skin irritation to
pitting and even the development of disfiguring scars. Accordingly, the
biophotonic materials and methods of the present disclosure can be used
to treat one or more of skin irritation, pitting, development of scars,
comedones, inflammatory papules, cysts, hyperkeratinazation, and
thickening and hardening of sebum associated with acne.

[0205] In certain embodiments, the biophotonic material of the present
disclosure is used in conjunction with systemic or topical antibiotic
treatment. For example, antibiotics used to treat acne include
tetracycline, erythromycin, minocycline, doxycycline, which may also be
used with the compositions and methods of the present disclosure. The use
of the biophotonic material can reduce the time needed for the antibiotic
treatment or reduce the dosage.

[0206] (iv) Wound Healing

[0207] The biophotonic materials and methods of the present disclosure may
be used to treat wounds, promote wound healing, promote tissue repair
and/or prevent or reduce cosmesis including improvement of motor function
(e.g. movement of joints). Wounds that may be treated by the biophotonic
materials and methods of the present disclosure include, for example,
injuries to the skin and subcutaneous tissue initiated in different ways
(e.g., pressure ulcers from extended bed rest, wounds induced by trauma
or surgery, burns, ulcers linked to diabetes or venous insufficiency,
wounds induced by conditions such as periodontitis) and with varying
characteristics. In certain embodiments, the present disclosure provides
biophotonic materials and methods for treating and/or promoting the
healing of, for example, burns, incisions, excisions, lesions,
lacerations, abrasions, puncture or penetrating wounds, surgical wounds,
contusions, hematomas, crushing injuries, amputations, sores and ulcers.

[0208] Biophotonic materials and methods of the present disclosure may be
used to treat and/or promote the healing of chronic cutaneous ulcers or
wounds, which are wounds that have failed to proceed through an orderly
and timely series of events to produce a durable structural, functional,
and cosmetic closure. The vast majority of chronic wounds can be
classified into three categories based on their etiology: pressure
ulcers, neuropathic (diabetic foot) ulcers and vascular (venous or
arterial) ulcers.

[0209] For example, the present disclosure provides biophotonic materials
and methods for treating and/or promoting healing of a diabetic ulcer.
Diabetic patients are prone to foot and other ulcerations due to both
neurologic and vascular complications. Peripheral neuropathy can cause
altered or complete loss of sensation in the foot and/or leg. Diabetic
patients with advanced neuropathy lose all ability for sharp-dull
discrimination. Any cuts or trauma to the foot may go completely
unnoticed for days or weeks in a patient with neuropathy. A patient with
advanced neuropathy loses the ability to sense a sustained pressure
insult, as a result, tissue ischemia and necrosis may occur leading to
for example, plantar ulcerations. Microvascular disease is one of the
significant complications for diabetics which may also lead to
ulcerations. In certain embodiments, biophotonic materials and methods of
treating a chronic wound are provided here in, where the chronic wound is
characterized by diabetic foot ulcers and/or ulcerations due to
neurologic and/or vascular complications of diabetes.

[0210] In other examples, the present disclosure provides biophotonic
materials and methods for treating and/or promoting healing of a pressure
ulcer. Pressure ulcers include bed sores, decubitus ulcers and ischial
tuberosity ulcers and can cause considerable pain and discomfort to a
patient. A pressure ulcer can occur as a result of a prolonged pressure
applied to the skin. Thus, pressure can be exerted on the skin of a
patient due to the weight or mass of an individual. A pressure ulcer can
develop when blood supply to an area of the skin is obstructed or cut off
for more than two or three hours. The affected skin area can turn red,
become painful and necrotic. If untreated, the skin can break open and
become infected. A pressure ulcer is therefore a skin ulcer that occurs
in an area of the skin that is under pressure from e.g. lying in bed,
sitting in a wheelchair, and/or wearing a cast for a prolonged period of
time. Pressure ulcers can occur when a person is bedridden, unconscious,
unable to sense pain, or immobile. Pressure ulcers often occur in honey
prominences of the body such as the buttocks area (on the sacrum or iliac
crest), or on the heels of foot.

[0211] Additional types of wounds that can be treated by the biophotonic
materials and methods of the present disclosure include those disclosed
by U.S. Pat. Appl. Publ. No. 20090220450, which is incorporated herein by
reference.

[0212] There are three distinct phases in the wound healing process.
First, in the inflammatory phase, which typically occurs from the moment
a wound occurs until the first two to five days, platelets aggregate to
deposit granules, promoting the deposit of fibrin and stimulating the
release of growth factors. Leukocytes migrate to the wound site and begin
to digest and transport debris away from the wound. During this
inflammatory phase, monocytes are also converted to macrophages, which
release growth factors for stimulating angiogenesis and the production of
fibroblasts.

[0213] Second, in the proliferative phase, which typically occurs from two
days to three weeks, granulation tissue forms, and epithelialization and
contraction begin. Fibroblasts, which are key cell types in this phase,
proliferate and synthesize collagen to fill the wound and provide a
strong matrix on which epithelial cells grow. As fibroblasts produce
collagen, vascularization extends from nearby vessels, resulting in
granulation tissue. Granulation tissue typically grows from the base of
the wound. Epithelialization involves the migration of epithelial cells
from the wound surfaces to seal the wound. Epithelial cells are driven by
the need to contact cells of like type and are guided by a network of
fibrin strands that function as a grid over which these cells migrate.
Contractile cells called myofibroblasts appear in wounds, and aid in
wound closure. These cells exhibit collagen synthesis and contractility,
and are common in granulating wounds.

[0214] Third, in the remodeling phase, the final phase of wound healing
which can take place from three weeks up to several years, collagen in
the scar undergoes repeated degradation and re-synthesis. During this
phase, the tensile strength of the newly formed skin increases.

[0215] However, as the rate of wound healing increases, there is often an
associated increase in scar formation. Scarring is a consequence of the
healing process in most adult animal and human tissues. Scar tissue is
not identical to the tissue which it replaces, as it is usually of
inferior functional quality. The types of scars include, but are not
limited to, atrophic, hypertrophic and keloidal scars, as well as scar
contractures. Atrophic scars are flat and depressed below the surrounding
skin as a valley or hole. Hypertrophic scars are elevated scars that
remain within the boundaries of the original lesion, and often contain
excessive collagen arranged in an abnormal pattern. Keloidal scars are
elevated scars that spread beyond the margins of the original wound and
invade the surrounding normal skin in a way that is site specific, and
often contain whorls of collagen arranged in an abnormal fashion.

[0216] In contrast, normal skin consists of collagen fibers arranged in a
basket-weave pattern, which contributes to both the strength and
elasticity of the dermis. Thus, to achieve a smoother wound healing
process, an approach is needed that not only stimulates collagen
production, but also does so in a way that reduces scar formation.

[0217] The biophotonic materials and methods of the present disclosure
promote the wound healing by promoting the formation of substantially
uniform epithelialization; promoting collagen synthesis; promoting
controlled contraction; and/or by reducing the formation of scar tissue.
In certain embodiments, the biophotonic materials and methods of the
present disclosure may promote wound healing by promoting the formation
of substantially uniform epithelialization. In some embodiments, the
biophotonic materials and methods of the present disclosure promote
collagen synthesis. In some other embodiments, the biophotonic materials
and methods of the present disclosure promote controlled contraction. In
certain embodiments, the biophotonic materials and methods of the present
disclosure promote wound healing, for example, by reducing the formation
of scar tissue.

[0218] In the methods of the present disclosure, the biophotonic materials
of the present disclosure may also be used in combination with negative
pressure assisted would closure devices and systems.

[0219] In certain embodiments, the biophotonic material is kept in place
for up to one, two or 3 weeks, and illuminated with light which may
include ambient light at various intervals. In this case, the composition
may be covered up in between exposure to light with an opaque material or
left exposed to light.

(6) Kits

[0220] The present disclosure also provides kits for preparing a
biophotonic material and/or providing any of the components required for
forming biophotonic materials of the present disclosure.

[0221] In some embodiments, the kit includes containers comprising the
components or compositions that can be used to make the biophotonic
materials of the present disclosure. In some embodiments, the kit
includes a biophotonic material of the present disclosure. The different
components making up the biophotonic materials of the present disclosure
may be provided in separate containers. For example, if the biophotonic
material is to include an oxygen-rich agent, the oxygen-rich agent is
preferably provided in a container separate from the chromophore.
Examples of such containers are dual chamber syringes, dual chamber
containers with removable partitions, sachets with pouches, and
multiple-compartment blister packs. Another example is one of the
components being provided in a syringe which can be injected into a
container of another component.

[0222] In other embodiments, the kit comprises a systemic drug for
augmenting the treatment of the biophotonic material of the present
disclosure. For example, the kit may include a systemic or topical
antibiotic, hormone treatment (e.g. for acne treatment or wound healing),
or a negative pressure device.

[0223] In certain embodiments, the kit comprises a first component
comprising a first chromophore; and a second component comprising at
least one thickening agent, wherein the thickening agent can form a
cohesive matrix when mixed with the first component, when the mixture is
applied to skin, or when illuminated with light.

[0224] In other embodiments, the kit comprises a means for applying the
components of the biophotonic materials.

[0225] In certain aspects, there is provided a container comprising a
chamber for holding a biophotonic material, and an outlet in
communication with the chamber for discharging the biophotonic material
from the container, wherein the biophotonic material comprises at least
one chromophore in a carrier medium which can form a biophotonic material
after being discharged from the sealed chamber, for example on contact
with skin or on illumination with a light. The container can be a
pressurized or non-pressurized spray can.

[0226] In certain embodiments, the kit comprises a first component
comprising the biophotonic material or a non-cohesive form of the
biophotonic material (`precursor`), and the second component comprises a
dressing or a mask. The dressing or mask may be a porous or semi-porous
structure for receiving the biophotonic material. The dressing or mask
may also comprise woven or non-woven fibrous materials. The biophotonic
material or its precursor can be incorporated, such as by injection, into
the dressing before the biophotonic material takes on a cohesive form
within the dressing or mask.

[0227] In certain embodiments of the kit, the kit may further comprise a
light source such as a portable light with a wavelength appropriate to
activate the chromophore the biophotonic material. The portable light may
be battery operated or re-chargeable.

[0228] Written instructions on how to use the biophotonic materials in
accordance with the present disclosure may be included in the kit, or may
be included on or associated with the containers comprising the
compositions or components making up the biophotonic materials of the
present disclosure. The instructions can include information on how to
form the cohesive matrix from the thickening agent(s) or matrix
precursors provided with the kit.

[0229] Identification of equivalent biophotonic materials, methods and
kits are well within the skill of the ordinary practitioner and would
require no more than routine experimentation, in light of the teachings
of the present disclosure.

[0230] Variations and modifications will occur to those of skill in the
art after reviewing this disclosure. The disclosed features may be
implemented, in any combination and subcombinations (including multiple
dependent combinations and subcombinations), with one or more other
features described herein. The various features described or illustrated
above, including any components thereof, may be combined or integrated in
other systems. Moreover, certain features may be omitted or not
implemented. Examples of changes, substitutions, and alterations are
ascertainable by one skilled in the art and could be made without
departing from the scope of the information disclosed herein. All
references cited herein are incorporated by reference in their entirety
and made part of this application.

[0231] Practice of the disclosure will be still more fully understood from
the following examples, which are presented herein for illustration only
and should not be construed as limiting the disclosure in any way.

EXAMPLES

Example 1

Preparation of an Exemplary Cohesive Biophotonic Material

[0232] A cohesive biophotonic material was prepared according to an
embodiment of the present disclosure and as summarized in Table 1.

[0233] Phase A was prepared by mixing water, eosin Y, rose bengal and
glucosamine sulphate. Phase B (water, glycerine, propylene glycol, urea
peroxide, carbopol) was then added to Phase A, and mixed until a light
viscous liquid was obtained. Phase C (sodium hyaluronate) was then added
to the mixture, and mixed until a homogenous thick cohesive gel was
obtained. This cohesive homogenous gel was spread onto a flat surface,
covered with an aluminum sheet and allowed to dry for 24 hours. After 24
hours, the resulting membrane was, easy to manipulate, and could be
applied to the skin and peeled off with little or no residue remaining. A
5-20% weight loss of the total weight of the material was found to occur
after drying for 24 hours. The membrane could be stored between two
layers of saran wrap, paraffin etc. On illumination with light (peak
wavelength between 400-470 nm and a power density of about 30-150
mW/cm2) for 5 minutes at a distance of 5 cm from the light source,
the film emitted fluorescent light which was captured by a
photospectrometer (SP-100 spectroradiometer (SP-100, ORB Optronix) to
measure the power density spectra versus wavelength and is illustrated in
FIG. 3. The emitted fluorescent light was in the green, yellow and orange
portions of the electromagnetic spectrum. An at least partial
photobleaching of the chromophores was observed after 5 minutes of
illumination.

Example 2

Angiogenic Potential of a Biophotonic Composition

[0234] The angiogenic potential of a biophotonic composition was evaluated
using a human skin model containing fibroblasts and keratinocytes. The
composition was a transparent gel comprising fluorescent chromophores,
eosin Y and erythrosine. Briefly, the biophotonic composition was placed
on top of the human skin model such that they were separated by a nylon
mesh of 20 micron pore size. The composition was then irradiated with
blue light (`activating light`) for 5 minutes at a distance of 10 cm from
the light source. The activating light consisted of light emitted from an
LED lamp having an average peak wavelength of about 400-470 nm and a
power density of about 30-150 mW/cm2. At a 10 cm distance from the
LEDs, the activating light had a power at the peak wavelength of about
2-3 mW/cm2/nm (about 2.5 mW/cm2/nm), an average power of about
55-65 mW/cm2, and a fluence in 5 minutes of irradiation of about
15-25 J/cm2 (about 16-20 J/cm2). Upon illumination with the
activating light, the biophotonic composition emitted fluorescent light,
as measured using a SP-100 spectroradiometer (SP-100, ORB Optronix) and
illustrated in FIG. 4. As the composition allowed the activating light to
pass therethrough, the skin model was illuminated substantially
simultaneously by both the activating light and the fluorescent light.

[0235] Since the biophotonic composition was in limited contact with the
cells, the fibroblasts and keratinocytes were exposed mainly to the
activating light and the fluorescent light emitted from the biophotonic
composition. Conditioned media from the treated human 3D skin model were
then applied to human aortic endothelial cells and diseased microvascular
endothelial cells from diabetic patients previously plated in
Matrigel®. The formation of tubes by endothelial cells was observed
and monitored by microscopy after 24 hours. The conditioned medium from
3D skin models treated with light illumination induced endothelial tube
formation in vitro, suggesting an indirect effect of the light treatment
(blue light and fluorescence) on angiogenesis via the production of
factors by fibroblasts and keratinocytes. Plain medium and conditioned
medium of untreated skin samples were used as a control, and did not
induce endothelial tube formation.

Example 3

Protein Secretion and Gene Expression Profiles of a Biophotonic
Composition

[0236] Wounded and unwounded 3D human skin models (EpiDermFT®, MatTek
Corporation) were used to assess the potential of a composition to
trigger distinct protein secretion and gene expression profiles. The
biophotonic composition comprised fluorescent chromophores eosin Y and
erythrosine. The composition was placed on top of wounded and unwounded
3D human skin models cultured under different conditions (with growth
factors, 50% growth factors and no growth factors). The skin models and
the composition were separated by a nylon mesh of 20 micron pore size.
Each skin model-composition combination was then irradiated with blue
light (`activating light`) for 2 minutes by light having a profile
similar to that described in Example 2. The fluorescence emission is
shown in FIG. 4. The controls consisted of 3D skin models not illuminated
with light.

[0237] Gene expression and protein secretion profiles were measured 24
hours post-light exposure. Cytokine secretion was analyzed by antibody
arrays (RayBio Human Cytokine antibody array), gene expression was
analyzed by PCR array (PAHS-013A, SABioscience) and cytotoxicity was
determined by GAPDH and LDH release. Results (Tables 2 and 3) showed that
the light treatment is capable of increasing the level of protein
secreted and gene expression involved in the early inflammatory phase of
wound healing in wounded skin inserts and in non-starvation conditions.
Interestingly, the effect of the light treatment on unwounded skin models
has a much lower impact at the cellular level than on wounded skin
insert, which suggests an effect at the cellular effect level of the
light treatment. It seems to modulate the mediators involved in
inflammation. Cytotoxicity was not observed in the light treatments.

[0238] The fluorescence spectra of biophotonic materials with different
concentrations of chromophores were investigated using a
spectrophotometer and an activating blue light. Exemplary fluorescence
spectra of Eosin Y and Fluorescein are presented in FIGS. 5a and 5b,
respectively. It was found that emitted fluorescence from the
chromophores increase rapidly with increasing concentration but slows
down to a plateau with further concentration increase. Activating light
passing through the composition decreases with increasing chromophore
composition as more light is absorbed by the chromophores. Therefore, the
concentration of chromophores in biophotonic materials of the present
disclosure can be selected according to a required ratio and level of
activating light and fluorescence treating the tissue based on this
example. The thickness of the biophotonic material can also be modulated
to control the light treating the tissues, as well as the optical
properties of the composition such as transparency.

Example 5

Synergistic combination of Eosin Y and Fluorescein

[0239] The photodynamic properties of (i) Fluorescein sodium salt at about
0.09 mg/mL, (ii) Eosin Y at about 0.305 mg/mL, and (iii) a mixture of
Fluorescein sodium salt at about 0.09 mg/mL and Eosin Y at about 0.305
mg/mL in a gel (comprising about 12% carbamide peroxide), were evaluated.
A flexstation 384 II spectrometer was used with the following parameters:
mode fluorescence, excitation 460 nm, emission spectra 465-750 nm. The
absorption and emission spectra are shown in FIGS. 6a and 6b,
respectively, which indicate an energy transfer between the chromophores
in the combination. It is to be reasonably inferred that this energy
transfer can also occur in biophotonic materials of the present
disclosure.

Example 6

Synergistic combination of Eosin Y Fluorescein and Rose Bengal

[0240] The photodynamic properties of (i) Rose Bengal at about 0.085
mg/mL, (ii) Fluorescein sodium salt at about 0.44 mg/mL final
concentration, (ii) Eosin Y at about 0.305 mg/mL, and (iii) a mixture of
(i), (ii) and (iii) in a gel (comprising about 12% carbamide peroxide)
(Set A), were evaluated. A flexstation 384 II spectrometer was used with
the following parameters: mode fluorescence, excitation 460 nm, emission
spectra 465-750 nm. The absorbance and emission spectra are shown in
FIGS. 7a and 7b, respectively, which indicate an energy transfer between
the chromophores in the chromophore combination. It is to be reasonably
inferred that this energy transfer can also occur in biophotonic
materials of the present disclosure.

[0241] Energy transfer was also seen between: Eosin Y and Rose Bengal;
Phloxine B and Eosin Y; Phloxine B, Eosin Y and Fluorescein, amongst
other combinations. It is to be reasonably inferred that energy transfer
can also occur in biophotonic materials of the present disclosure.

Example 7

Collagen Formation Potential of a Biophotonic Composition

[0242] A biophotonic composition comprising 0.01% eosin Y and 0.01%
fluorescein in a carrier matrix (1.8% carbopol gel) was evaluated for its
potential to induce collagen formation. Dermal human fibroblasts were
plated in glass-bottomed dishes with wells (MatTek®). There were
approximately 4000 cells per well. After 48 hours, the glass-bottomed
dishes were inverted and the cells were treated through the glass bottom
with (i) no light (control), (ii) sunlight exposure for about 13 minutes
at noon (control), (iii) the composition applied to the glass well bottom
on the other side of the cells (no light exposure), (iv) the composition
applied to the glass well bottom on the other side of the cells and
exposed to sunlight for about 13 minutes at noon, and (v) the composition
applied to the glass well bottom on the other side of the cells and
illuminated with blue light. In the case of (iii), (iv) and v), there was
no direct contact between the cells and the composition. In the case of
(iv), the cells were exposed to emitted light from and through the Eosin
Y and Fluorescein composition when exposed to sunlight. A partial
photobleaching was observed in (iv) and total photobleaching in (v).
After the treatment, the cells were washed and incubated in regular
medium for 48 hours. A collagen assay was then performed on the
supernatant using the Piero-Sirius red method. This involved adding
Sirius red dye solution in picric acid to the supernatant, incubating
with gentle agitation for 30 minutes followed by centrifugation to form a
pellet. The pellet was washed first with 0.1N HCl and then 0.5 N NaOH to
remove free dye. After centrifugation, the suspension was read at 540 nm
for collagen type I. The results are shown in Table 4.

TABLE-US-00004
TABLE 4
A qualitative comparison of collagen type I concentration in a dermal
human fibroblast supernatant exposed to (i) no light (control), (ii)
sunlight
exposure for about 13 minutes at noon (control), (iii) any light emitted
from the Eosin Y and Fluorescein composition through a glass separation
(no activating light exposure), (iv) any light emitted from and through
the
Eosin Y and Fluorescein composition through a glass separation when
illuminated with sunlight exposure for about 13 minutes at noon, and
(v) light emitted from and through the composition through a
glass separation when illuminated with blue light.
Sunlight Eosin Y and Eosin Y and Eosin Y and
No light alone Fluorescein- Fluorescein- Fluorescein-
(control) (alone) no light sunlight blue light
Collagen + + ++ +++ +++
formation
++ indicates collagen levels about twice as high as +,
+++ indicates collagen levels about three times as high as +.

[0243] There was a statistical difference between the collagen levels
induced by the Eosin Y and Fluorescein composition exposed to sunlight
compared to the no light and sunlight alone controls. There was also a
statistical difference between the collagen levels induced by composition
exposed to blue light compared to the no light and sunlight alone
controls. Collagen generation is indicative of a potential for tissue
repair including stabilization of granulation tissue and decreasing of
wound size. It is also linked to reduction of fine lines, a decrease in
pore size, improvement of texture and improvement of tensile strength of
intact skin.

[0244] It is to be reasonably expected that the same or similar
biophotonic effects can be obtained with a cohesive biophotonic material
of the present disclosure providing substantially similar or equivalent
light emission properties as the compositions described in Examples 2, 3
and 7.

Example 8

Preparation of an Exemplary Cohesive Biophotonic Material Based on
Silicone

[0245] Cohesive biophotonic membranes were made, according to embodiments
of the present disclosure, comprising a silicone membrane having
incorporated therein chromophores, specifically water soluble
chromophores Eosin Y and Fluorescein. The biophotonic membranes were
based on a colloidal system comprising an aqueous phase of solubilized
chromophores within a solid silicone phase (micro-emulsion). The cohesive
biophotonic membrane was made by mixing a base (B) comprising (1)
dimethyl siloxane, dimethylvinyl terminated, (ii) dimethylvinylated and
trimethylated silica, and (iii) tetra(trimethoxysiloxy) silane in ethyl
benzene and with a curing agent (C) comprising (i) dimethyl,
methylhydrogen siloxane, (ii) dimethyl siloxane, dimethylvinyl
terminated, (iii) dimethylvinylated and trimethylated silica, and (iv)
tetramethyl tetravinyl cyclotetra siloxane in ethyl benzene (both in
liquid form from a Sylgard® 184 silicone elastomer kit, Dow Corning
Corp, Ltd). When mixed at a ratio of 10 (B): 1 (C), the mixture cures to
an elastic material. The material obtained was a flexible and
transparent/translucent elastomer. A stabilizing agent was also used to
stabilize the emulsion and avoid phase separation. In one example,
carboxymethyl cellulose (CMC) was used as the stabilizing agent (about
2%). In another example, gelatin was used as the stabilizing agent.

[0246] In one embodiment, 9.4 g of the base was mixed with 0.94 g of the
curing agent, and to this was added 2 mL of 2% CMC solution (18 wt %)
containing 0.327 mg (0.011 wt % within the aqueous phase) of eosin Y and
0.327 mg (0.011 wt % within the aqueous phase) of fluorescein. The whole
mixture was emulsified vigorously for about 15 minutes and cast on a
petri dish for curing at 35° C. for about 16 hours forming a
translucent/transparent membrane comprising a silicone matrix with
embedded droplets of the chromophore in CMC phase. In another embodiment,
2 mL of gelatin solution (5%) was used as the stabilizing agent instead
of CMC. This also formed a translucent/transparent membrane comprising a
silicone matrix with embedded droplets of the chromophores in the gelatin
phase. In both cases, a 2 mm thick membrane was achieved, although it
will be understood that the thickness of the membrane can be controlled
by the volume of cast solution. In both cases, the membranes could be
applied and removed from tissue (human skin) in one piece.

[0247] It will be appreciated that other stabilizing agents which can be
used which include but are not limited to methyl cellulose or
hydroxyethylcellulose. Other concentrations of gelatin can be used such
as from about 1 to about 20 wt %. The total weight percent of the aqueous
phase can range from about 2 weight % to about 40 weight %.

[0248] When the biophotonic membranes were illuminated with blue light,
the chromophores absorbed and emitted light. An at least partial
photobleaching of the chromophores was observed with time of
illumination. When the water soluble fluorescent chromophores were
incorporated directly into the silicone (i.e. as a single phase), they
did not absorb or emit light. It is believed by the inventors that their
inclusion in the silicone membrane as an aqueous phase provided the
appropriate medium to allow biophotonic activity. Instead of a liquid
phase, the water soluble chromophores could also be directly surrounded
by any other medium which allows the absorption and emission of light,
such as a gel or water, or adsorbed on fine solid particles such as, but
not limited to, silica and hydroxyapatite particles.

[0249] The above example can also be demonstrated using any other
liposoluble polymers or matrices, instead of silicone.

Example 9

Preparation of an Exemplary Cohesive Biophotonic Material Based on Gelatin

[0250] A cohesive biophotonic material was made, according to another
embodiment of the present disclosure, comprising a cohesive gelatin
matrix incorporating therein chromophores. In a typical preparation, 10 g
of gelatin was dispersed in 50 mL of de-ionized water then heated to
around 65° C. in a hot water bath under continuous stirring until
complete dissolution of gelatin. While the temperature was decreased to
around 40° C., 0.5 mL of eosin Y solution (10.9 mg/mL) was added
to the gelatin solution, and the resulting gelatin solution (20% w/v)
including eosin Y was cast on a petridish and cooled down to room
temperature to form a hydrogel membrane of gelatin containing eosin Y. A
transparent elastic membrane of 2 mm was obtained. The membrane could be
applied and removed from tissue in one piece. When the gelatin membrane
was illuminated with blue light, the chromophore absorbed and emitted
light. An at least partial photobleaching of the chromophore within the
cohesive membrane was observed after illumination. A similarly peelable
membrane was also obtained with a gelatin matrices having more than 5 wt
%. Peelable biophotonic membranes having <about 5 weight % gelatin
could be obtained by adding chemical cross-linkers such as glutaraldehyde
or glyoxal. Similar results were also obtained using chitosan as the
cohesive matrix instead of gelatin.

Example 10

Measurement of Tensile Strength

[0251] The tensile strength of certain embodiments of the silicone and
gelatin-based cohesive biophotonic materials formed according to Examples
8 and 9 were measured according to the following method. Rectangular test
samples of 50 mm×10 mm having a 2 mm thickness were prepared based
on the silicone and gelatin membranes of Examples 8 and 9 as well the
membranes without chromophore(s). Sample length, width and thickness were
verified at 3 points per dimension using a Vernier caliper and were used
to calculate the cross-section area of the samples.

[0252] Each end of the sample was tightly fixed between a clamp with a 15
mm rubber grip linked to a 1/16'' steel cable. This sample/clamp assembly
was installed vertically in a rigid scaffold made of steel tubes. The top
cable was hung from a manual ratcheting device for winching the top cable
away from the bottom cable, and the bottom cable was attached to a
weight. The weight was loaded on a precision balance which was installed
vertically under the manual ratcheting device. The sample between the
clamps was then stretched at a steady slow rate using the winch. The
force required to deform the sample was measured by the decrease of
weight measured on the balance relative to a baseline length. The
baseline was measured by relaxing the sample so that the weight measured
by the balance was maximal. The top cable was then pulled away from the
bottom cable via the ratcheting mechanism until a weight decrease was
observed on the scale. This point was considered baseline and the reading
on the balance was recorded and the length of the sample (distance
between the clamps) was measured with a Vernier caliper. This length was
defined as the initial length of the sample. The ratchet was then
activated stepwise to stretch the sample with the balance reading and
sample length being recorded at every step until rupture of the sample.
Absence of grip slippage was verified by checking the stabilization of
the measured weight and using visual indicators on the samples.

[0253] Typical stress-strain curves for the silicone-based and the
gelatin-based membranes are shown in FIGS. 8a and 8b, respectively. The
silicone membranes with and without chromophores, and with different
thickening agents, had substantially similar tensile properties. The
gelatin membranes with and without chromophores also had substantially
similar tensile properties. The gelatin-based membranes had a tensile
strength of about 0.01 MPa (±10%) (100 kPa) and an Elastic Modulus
(slope of the stress/strain curve) of about 0.01 MPa (±10%) (100 kPa).
The silicone-based membranes were stiffer than the gelatin-based
membranes and had an average Elastic Modulus of about 1.11 MPa (±10%)
(1110 kPa). This was well within the range reported in literature of
about 1.2-1.8 MPa) The measured tensile strength was 0.405 MPa (826 g)
due to grip slippage but is expected to be up to about 8 MPa based on
literature reports on cured silicone.

[0254] This methodology was based on a similar principle of operation as
American Society for Testing and Materials tensile testing methods such
as ASTM D638, ASTM D882 and ASTM D412. However, instead of a pneumatic
force, in the present example, gravity was used for sample extension.

Example 11

Measurement of Adhesion Strength

[0255] The adhesion strength of certain embodiments of the biophotonic
materials formed according to Examples 8 and 9 were measured according to
the following method. Samples were prepared as described in Example 10.
One end of each sample was fixed to a clamp with a 15 mm rubber grip
linked to one end of a 1/16'' steel cable. The other end of the cable,
via a low-friction pulley, was attached to a weight placed on a balance.
The sample was laid flat on the skin of an inside forearm of a volunteer.
A known weight, of surface area matching the sample, was then placed on
the sample in order to apply a homogenous and known downwards force on
the sample contacting the skin. The normal force Fn (force exerted
by each surface on the other in a perpendicular direction to the surface)
was calculated by multiplying the combined weight of the sample and the
weight on the sample by the gravity constant, g (9.8 m/s2). The
forearm, with the sample loaded with the weight, was then pulled away
from the cable until the sample slipped from the skin surface. The weight
recorded on the balance at this time was calculated by multiplying g to
obtain the force of friction (Fr) (force required to overcome the
friction between the sample and the skin). The friction coefficient of
the sample can then be calculated using Fr≦μFn
(Coulomb's friction law).

[0256] On average, the silicone-based membranes had a friction coefficient
of about 1.43, and the gelatin-based membranes had a friction coefficient
of about 1.04. These values can be converted to the weight required to
shear off a sample from the test surface by multiplying the friction
coefficient by the sample weight. So, for the silicone-based membranes, a
weight of 1.50 g is required to shear-off the membranes from skin. From
FIG. 8a, this is equivalent to an elongation of about 0.1% and is well
below its tensile strength. For the gelatin-based membranes, a weight of
about 1.04 g was required to shear-off the membranes from skin. From FIG.
8b, this is equivalent to an elongation of about 1.5% and is well below
its tensile strength (equivalent to 24.12 g). Therefore, all the
silicone-based membranes and gelatin-based membranes of Examples 8 and 9
were peelable.

Example 12

Demonstration of Peelable Nature of Cohesive Biophotonic Materials of the
Present Disclosure

[0257] The biophotonic materials described in Examples 1, 8 and 9 were
evaluated for peelability by applying them to the skin of volunteers and
peeling off by hand. All membranes could be peeled off, reapplied and
peeled off again without damage to the membranes and without leaving
residues on the volunteer skins.

Example 13

Cell Studies

[0258] Certain embodiments of the cohesive biophotonic materials of
Example 8 were evaluated for their ability to modulate inflammation,
specifically cytokines IL6 and IL8. HaCaT cells were used as an accepted
in vitro module for assessing modulation of these inflammatory cytokines.
A non-toxic concentration of IFNγ was used to modulate the
secretion of IL6 and IL8 by the HaCaT cells.

[0259] Silicone membranes containing an aqueous phase of eosin y and
fluorescein and including either CMC or gelatin in the aqueous phase were
evaluated. The anti-inflammatory effect of Dexamethasone was used as a
positive control at a concentration of 5 μM. The materials were
illuminated with blue light for 90 seconds at a distance of 5 cm at a
fluence of about 11.5 J/cm2. Cytokine quantification was performed
by cytokine ELISA on the culture supernatant 24 hours after treatment.
The quantity of cytokine secreted was normalized to cell viability. No
toxic effect was observed for all the test samples as measured by cell
viability using a spectrophotometric evaluation of viable cell number 24
hours after treatment. All of the membranes tested, produced a downward
modulation of IL6 and IL8 on IFNγ stimulated HaCaT cells.

[0260] It should be appreciated that the invention is not limited to the
particular embodiments described and illustrated herein but includes all
modifications and variations falling within the scope of the invention as
defined in the appended claims.